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i
CM
C REVIEW M EMORANDUM
Date: AUGUST 2 1, 2021
To: The Biologics License Application (BLA) File STN 125742
From: Xiao Wang, OVRR/ DVP, CMC Product Reviewer
Through: Anissa Cheung, OVRR/DVP
Keith Peden, Lab Chief, OVRR/DVP
Robin Levis, Deputy Director, OVRR/DVP
Jerry Weir, Director, OVRR/DVP
CC: Ramachandra Naik, Chair, OVRR/DVRPA
Capt. Michael Smith, RPM, OVRR/DVRPA
Laura Gottschalk, RPM, OVRR/DVRPA
Applicant : BioNTech Manufacturing GmbH (in partnership with Pfizer Inc.)
Subject: CMC Review of Original BLA STN 125742.0 ;
Product : Pfizer -BioNTech COVID- 19 Vaccine; Human Coronavirus mRNA Vaccine
for the Prevention of Coronavirus Disease 2019 (COVID -19)
ii
The following abbreviations are used throughout the memorandum:
AM
TE A nalytical Method Transfer Exercise
ATM Animal Tria l Material
BLA Biologics License Application
BNT BioNTech Manufacturing GmbH
CM
C Chemistry, Manufacturing, and Control
CoA Certificate of Analysis
CPP Critical Process P arameter
CQA Critical Quality Attribute
CTM Clinical Trial Material
dLIA Direct Luminex Assay
DS Drug Substance
DP Drug Product
EUA Emergency Use Authorization
FC Fi
nal Container
GMP Good Manufacturing Practice
IM Intramuscular
IPT-C In- process Tests for C ontrol
IPT-M In- process Tests for Monitoring
IR Information Request
L
NP Lipid N anoparticle
LPQ Laboratory Process Qualification
MCB Master Cell Bank
modRNA N ucleoside -modified Messenger RNA
NHP Nonhuman Primate
PAI Pre-approval Inspection
PPQ Process Performance Qualification
RBD Receptor Binding Domain
RH Relative Humidity
RPH Relative Process History
W
CB Working Cell Bank
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able of Contents
1.Product Name/Proprietary Name/Product Type........................................................ 1
2.Submissions reviewed ................................ .............................................................. 1
3.Executive Summary and Recommendation ................................ .............................. 2
4.BNT162b2 Drug Substance ................................ ...................................................... 5
4.1 General Description ............................................................................................... 5
4.2 Manufacturers ........................................................................................................ 5
4.3 Control of M aterials ................................................................................................ 6
4.4 Description of Drug Substance Manufacturing Process and Process Controls .... 11
4.5 Process Validation for Drug Substance Manufacturing ........................................ 15
4.6 DS Manufacturing Process Development ............................................................ 23
4.7 Impurity Profile of the Drug Substance ................................................................. 26
4.8 Adventitious Agents Safety Evaluation ................................................................ 28
4.9 Characterization Studies on Drug Substance ...................................................... 28
4.10 Control of Drug Substance ................................................................................. 30
4.11 Analytical Procedures for Drug Substance ......................................................... 33
4.12 Validation of Analytical Procedures .................................................................... 35
4.13 Reference Standards ......................................................................................... 42
4.14 Summary of Batch Analysis ............................................................................... 43
4.15 Container -Closure System ................................................................................. 43
4.16 Stability of Drug Substance ................................................................................ 44
5. BNT162b2 Drug Product ........................................................................................... 49
5.1 General Description and Composition .................................................................. 49
5.2 Manufacturers ...................................................................................................... 50
5.3 Control of Excipients ............................................................................................ 51
5.4 Description of Drug Product Manufacturing Process and Process Controls ........ 59
5.5 Process Validation ............................................................................................... 65
5.6 DP Manufacturing Process Development ............................................................ 80
5.7 Impurity Profile of the Drug Product ..................................................................... 83
5.8 Characterization Studies on Drug Product ........................................................... 84
5.9 Control of Drug Product ....................................................................................... 84
5.10 Analytical Procedures for Drug Product ............................................................. 90
iv 5.
11 Validation of Analytical Procedures .................................................................... 93
5.12 Reference Standards ....................................................................................... 100
5.13 Summary of Batch Analysis ............................................................................. 101
5.14 Container -Closure System of the Final Container ............................................ 101
5.15 Stability of Drug Product .................................................................................. 102
6.Other CMC -r elated Information Not Covered in Module 3 .................................... 108
7.Follow- up on Ongoing Issues Identified at the Time of EUA Request .................. 109
8.Pre-A pproval Inspection ....................................................................................... 112
9.Nonclinical Studies ................................ ............................................................... 112
9.1 Nonclinical Pharmacology Studies ..................................................................... 112
9.2 Nonclinical Pharmacokinetics (PK) Evaluation ................................................... 115
10. Clinical Assays ................................ .................................................................. 116
10.1 Diagnostic Assays Used to Support Clinical Efficacy Endpoints ...................... 116
10.2 Immunogenicity Assays Used for Exploratory Immunogenicity Endpoints ....... 118
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1.Product Name/P roprietary N ame/P roduct Type
Product: Pfizer -BioNTech COVID- 19 Vaccine
Proprietary name: COMIRNATY
Nonproprietary name: COVID- 19 Vaccine , mRNA
Product Type: Human Coronavirus mRNA vaccine expressing SARS- CoV- 2 spike
glyco protein (BioNTech code number BNT162b2, Pfizer code number PF- 07302048);
the mRNA ( variant RBP020.2) is formulated with lipid s ALC- 0315, ALC -0159, DSPC ,
and cholesterol to form lipid nanoparticles (LNPs) .
2.Submissions reviewed
Date Received Submission Contents/Comments
May 6 , 2021 STN 125742/0 BLA R oll 1 submission including nonclinical
pharmacology and pharmacokinetics studies and
clinical assays
May 18, 2021 STN 125742/ 0.1 BLA Roll 2 submission encompassing all quality -
related information in Module 3
July 9, 2021 STN 125742/ 0.10 Lot-release protocol template for the drug product
including the assay performed and acceptance c riteria
July 15, 2021 STN 125742/ 0.11 Request for an exception to the 21 CFR 610.15(a) for
the vaccine as a preservative -free presentation
July 23, 2021 STN 125742/ 0.16 Response to DVP/DBSQC IR regarding the validation
of RNA testing by
July 28, 2021 STN 125742/ 0.19 Response to DVP IR regarding multiple CMC -related
issues (DS and DP manufacturing process and testing,
and clinical assays) and the categorical exclusion for
an environment analysis
August 2, 2021 STN 125742/ 0.27 CMC -related information described in the dra ft
pac
kage inser t
August 5, 2021 STN 125742 /0.31 Response to DVP/Stat istical IR regarding the validation
studies for the direct Luminex assay (dLIA) for IgG antibody quanti fication in human sera
August 6, 2021 STN 125742/ 0.33 Response to DVP IR regarding manufacturing process
validation issues
August 6, 2021 STN 125742/ 0.34 Response to DVP/Statistical IR regarding the validation
report (VAL100147509) for of the
vaccine DP by
August 9, 2021 STN 125742/ 0.35 Response to DBSQC/DVP IR regarding validation of the assay methods and lot release
August 9, 2021 STN 125742/ 0.36 Response to DMPQ IR regarding the saline diluent
August 10, 2021 STN 125742/ 0.39 Response to DVP IR regarding a step executed during the process at Pfizer Puurs
August 13, 2021 STN 125742/ 0.47 Response to DMPQ IR regarding the saline diluent
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Date Received Submission Contents/Comments
August 17, 2021 STN 125742/0.55 Stability data to support a shelf-life extension of the
undiluted DP up to 9 months when stored at the intended long-term storage condition
August 19, 2021 STN 125742/0.61 Response to DVP IR regarding the final commercial shelf life of the BNT162b2 DP and date of manufacture
August 19, 2021 STN 125742/0.62 Response to DVP IR regarding the final
3.Executive Summary and Recommendation
The BNT162b2 COVID -19 vaccine, developed under a collaborative agreement
between Pfizer and BioNTech (BNT), is a nucleoside- modified messenger RNA
(modRNA) -based vaccine candidate indicated for active immunization for the prevention
of coronavirus disease 2019 (COVID -19). The single- stranded mRNA encodes the
SARS- CoV-2 full -length spike (S) glycoprotein, which is codon- optimized and modified
to express the P2 mutant, a pre- fusion S protein (P2 S; version 9). The modRNA is
stabilized by formulation with lipids consisting of DSPC, cholesterol, ALC -0315, and
ALC- 0159 to generate lipid nanoparticles ( LNPs ). Other ingredients in the BNT162b2
vaccine include potassium chloride, monobasic potassium phosphate, sodium chloride,
dibasic sodium phosphate dihydrate, and sucrose.
The final vaccine product is a white to off -white, sterile, preservative -free, multi -dose
frozen suspension to be diluted with 0.9% sodium chloride injection, USP, for
intramuscular (IM) injection. The vaccine is filled in a 2 mL clear glass vial with a rubber
stopper (not made of natural rubber latex) as a multi -dose concentrate (after dilution
each vial contains 6 doses of 0.3 mL) and is administered as a series of two IM
immunizations with each dose containing 30 µg of modRNA. The second dose is
administered 21 days after the first dose. The undiluted vaccine vials should be stored
frozen between - 90°C to - 60°C and they may be stored at - 25°C to - 15°C for up to two
weeks. To facilitate storage and administration of the BNT162b2 vaccine at the point of
use, the thawed vials can be stored between 2°C to 8°C for up to 1 month (31 days)
prior to dilution; the diluted vaccine must be stored between 2°C to 25°C and used within 6 hours from the time of dilution. The vaccine is indicated for use in individuals 16 years of age or older.
The manufacturing process for the BNT162b2 drug substance (DS) consists of two major steps: . Two manufacturing
facilities (Pfizer ACMF and Pfizer Suite ) located in Andover, MA are involved in the commercial production of the BNT162b2 DS. The DS manufacturing process was
validated at both facilities by process -performance qualification (PPQ) studies ( and
PPQ lots manufactured at Pfizer ACMF and a t Pfizer Suite , respectively ). Consistency
of the DS manufacturing process was demonstrated by maintaining process parameters within defined ranges and obtaining satisfactory in- process and release testing result s.
Additionally, data obtained from the analytical comparability assessment for the ACMF
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and Suite DS batches further support the DS manufacturing process for the consistent
production of the BNT162b2 DS with acceptable quality.
The manufacturing process for the BNT162b2 drug product (DP) involves the
modRNA DS with lipid excipients during LNP formulation followed by fill and finish . The
two facilities proposed for the manufacture of the BNT162b2 DP are Pfizer Puurs
(located in Puurs, Belgium) and Pfizer Kalamazoo (located in Kalamazoo, Michigan,
USA) , both of which have been authorized for the manufacture of the BNT162b2 DP
under the Emergency Use Authorization (EUA 27034). Validation of the DP manufacturing process took place in two Phases. The Phase I network PPQ study ,
which involved multiple authorized DP manufacturing facilities under the EUA, was
executed to demonstrate that the manufacturing process consistently produces DP lots of acceptable quality across multiple supply nodes. The Phase II process validation occurred at the proposed commercial facilities with batches at and
batches at batch sizes being manufactured at Pfizer Puurs and
batches at and batches at batch sizes being manufactured at Pfizer Kalamazoo. Both Phase I and Phase II process -validation studies were executed
successfully according to pre- established protocols, thereby demonstrating that the DP
manufacturing process is consistent and well -controlled. Additionall y, a batch size
for the manufacture of BNT162b2 DP was validated at the Pfizer Puurs site. Analytical comparison of the process -validation batches to clinical and emergency supply lot data
further demonstrated comparable product quality from clinical through commercial
supply of the BNT162b2 DP. Overall, the process validation data support the capability
of the commercial manufacturing process to produce a consistent DP with acceptable quality at the proposed commercial manufacturing facilities.
Stability studies have been generated for both the DS and DP lots to support the
licensure of the BNT162b2 vaccine. All available stability data generated to date support
the initial commercial shelf life of for the BNT162b2 DS when stored at the
intended storage condition of For DP shelf -life determination, up to
months of stability data are available for one clinical batch at the intended storage
condition of - 70 ± 10°C and up to 9 months of stability data are available for
emergency supply DP lots when stored at the intended storage condition of - 90 to -
60°C . As comparable product quality has been demonstrated from clinical through
commercial supply of the BNT162b2 DP, the stability data from both the clinical trial materials (CTMs) and the emergency supply lot s are considered supportive of the
proposed shelf life of 9 months for the commercial BNT162b2 DP . The final commercial
shelf life of the BNT162b2 DP is intended to be months when stored at the intended storage condition of - 90 to - 60°C ; stability studies are ongoing. The available stability
data from the emergency supply/PPQ DP lots will be submitted post licensure at
intervals (9, months) as data become available to support future shelf -life
extensions .
The analytical procedures developed and used for the release and stability monitoring of BNT162b2 DS and DP include tests to ensure their identity, purity, quality, and
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potency. The assay methods are appropriately established and performed routinely
according to the established standard operating procedures ( SOPs) . Validation of each
assay method w as performed at all the proposed testing sites (either through
validation/co- validation or by analytical method transfer exercise) and the results have
demonstrated that the assay methods are accurate, specific, robust, and precise over
the specified range, i ndicating that they are suitable for the intended use.
Two notable issues were identified during the previous EUA 27034 review and were
resolved. The first issue involves t he occurrence of in
DP lots produced for emergency use, and the second issue involves the occasional
observation of visible intrinsic particles detected during visual inspection of filled DP
vials. Investigation of the identified that the are
only associated with the batches that were manufactured with a
operation at (a supplier of the ). Since the
implementation of a process at , the resulting DP lots
have been consistently demonstrating . Regarding the
intrinsic particles, the frequency of occurrence is low and DP vials containing intrinsic
particles can be detected and discarded through 100% automated/manual visual
inspection. The intrinsic particles consist of components used for
(i.e., they are not foreign particles) and the available data suggest that they
have minimal potential to impact product safety and quality.
Two clinical diagnostic assays (Cepheid Xpert Xpress RT- PCR assay for the detection
of SARS- CoV-2 in clinical specimens and Roche Elecsys Anti -SARS- CoV- 2 assay for
the evaluation of serostatus to SARS -CoV- 2) were used to assess clinical end points.
Both assays have received FDA authorization under EUA. Validation of both assays has
been performed at Pfizer’s testing facility (Pfizer Vaccine Research and Development;
Pearl River, NY) , and the results support the suitability of both assays for their intended
use in clinical studies .
Final Recommendation:
I recommend approval of this BLA .
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4.BNT162b2 Drug Substance
4.1 General Description
The BNT162b2 drug substance (DS) is a single- stranded, 5’ -capped mRNA encoding
the full -length SARS- CoV- 2 spike glycoprotein (S1S2 protein) derived from the Wuhan-
Hu-1 isolate (GenBank MN908947.3 and GenBank QHD43416.1). The antigen- coding
RNA sequence is codon- optimized and contains two proline mutations (
), which ensure expression of an antigenically optimal trimerized pre- fusion
confirmation (P2 S). The RNA also contains common structural elements, including 5’ -
cap, 5’ -UTR, 3’- UTR, and poly(A) tail, all of which are designed for mediating high RNA
stability and translation efficiency. is replaced with the
during the RNA transcription. This nucleoside substitution has
been demonstrated to enhance translation of in vitro transcribed mRNA while reducing
its reactogenicity .
The final BNT162b2 DS is a clear to
and is formulated at a target concentration of in DS
.
4.2 Manufacturers
Facilities and manufacturing sites involved in the commercial DS manufacturing and
testing are presented in Table 1.
Table 1. Sites and Responsibilities for Manufacture and Testing of BNT162b2 DS
Site FEI/DUNS Numbers Responsibility
(Pfizer, Andover, ACMF) (Pfizer,
Andover Building
and Building ) Wyeth BioPharma Division of Wyeth Pharmaceuticals, LLC
a
1 Burtt Road Andover, MA 01810
United States FEI: 1222181
DUNS: 174350868 ACMF /Suite : Manufacture
of drug substance
Bldg. : Release and Stability Testing
Pfizer Inc
875 Chesterfield Parkway West
Chesterfield, MO 63017
United States FEI: 1940118
DUNS: 004954111 Release and Stability
Testing
a.The legal entity name was changed at the acquisition by Pfizer in 2009; since then, the Wyet h
P
harmaceuticals manufacturing site in Andover, Massachusetts belongs to Pfizer’s production sites and is
embedded in Pfizer´s GMP system.
Reviewer’s Comments:
Two m anufacturing nodes were employed for DS manufacture under the initial EUA
27034 request: (1) Pfizer, Andover, ACMF and (2) BioNTech, Mainz, and
Germany. Pfizer, Andover, Suite was introduced as an
additional DS manufac turing site under the EUA 27034. Both Andover ACMF and
Suite manufacturing facilities are included in the BLA submission. The
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BNT/ manufacturing node is not intended for the manufacture of
commercial DS.
4.3 Control of Materials
Control of Non -Compendial Starting Materials and Raw Materials
Raw materials used in the DS manufacturing process are purchased from approved suppliers and are tested and released upon receipt in accordance with the applicant’s
internal quality control program.
The current acceptance criteria for non- compendial starting materials and raw materials
used during the manufacture of the BNT162b2 DS are presented in Table 2 below. Note
that starting materials are defined as a reagent or material used during the manufacture
of the BNT162b2 vaccine product that is intended to be part of the final product, such as
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4.8 Adventitious Agents Safety Evaluation
The adventitious agent control program includes the engineering systems of the facility
and vessels, the control of raw materials, various filtration steps to control microbial
burden in buffers and the process stream, and in- process and environmental testing to
monitor the level of adventitious agents in and around the process stream.
All raw materials used in the production of DS are evaluated as part of a comprehensi ve
program to identify and manage transmissible spongiform encephalopathy (TSE) /
bovine spongiform encephalopathy (BSE) risks. The only raw material of direct animal
origin was identified to be
Based on the comprehensive adventitious -agent control
program, all raw materials were found to be compliant with the EMA Note for Guidance (EMA/410/01 rev.3) and associated with minimal risk for TSE/BSE.
Other materials of animal origin may be used in the production of polymer for filters, manifolds, containers, and/or filter components. These equipment components may
contain traces of animal tallow derivatives. The tallow is processed under rigorous
conditions and is considered compliant with the TSE note for guidance (EMA/410/01).
4.9 Characterization Studies on Drug Substance
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5. BNT162b2 Drug Product
5.1 General Description and Composition
The BNT162b2 DP is a sterile dispersion of RNA -containing lipid nanoparticles (LNPs)
in aqueous cryoprotectant buffer. It is filled into multi -dose vials containing 0.45 mL of
the DP at . After dilution with 1.8 mL of sterile 0.9% sodium chloride solution,
each vial contains a total of six 0.3 mL doses, with each dose containing 30 µg of RNA.
The composition of DP, including t he BNT162b2 DS, lipid excipients, buffer, and
cryoprotectant, are listed in Table 22 below. The concentration and amount of each
component per DP container, and the corresponding amount per dose are also listed.
Table 22 . Composition of BNT162b2 Drug Product, Multi -dose Vial
Name of Ingredients Reference to
Standard Function Concentration
(mg/mL) Amount
per vial Amount
per dose
BNT162b2 DS In-house
specification Active ingredient 0.5 225 µg 30 µg
ALC-0315 In-house
specification Functional lipid 7.17 3.23 mg 0.43 mg
ALC-0159 In-house
specification Functional lipid 0.89 0.4 mg 0.05 mg
DSPC In-house
specification Structural lipid 1.56 0.7 mg 0.09 mg
Cholesterol Ph. Eur. and /or
USP-NF Structural lipid 3.1 1.4 mg 0.2 mg
Sucrose USP-NF, Ph. Eur. Cryoprotectant 103 46 mg 6 mg
Sodium chloride USP-NF, Ph. Eur. Buffer component 6 2.7 mg 0.36 mgc
Potassium chloride USP-NF, Ph. Eur.a Buffer component 0.15 0.07 mg 0.01 mg
Dibasic sodium
phosphate, dihydrate USP-NF, Ph. Eur. Buffer component 1.08 0.49 mg 0.07 mg
Monobasic potassium
phosphate USP-NF, Ph. Eur.a Buffer component 0.15 0.07 mg 0.01 mg
Water for injection USP-NF, Ph. Eur. Solvent/vehicle q.s.bq.s.bq.s.b
a.Supplier Certificate of Analysis confirms compliance to both USP -NF and Ph. Eur.; however,
incoming testing may be performed only in accordance with a site’s local compendia.
b.q.s. = quantum satis (as much as may suffice)
c.The diluent (0.9% sodium chloride Injection) contributes an additional 2.16 mg per dose
The four lipids used to encase the modRNA include:
•ALC- 0315: ((4-hydroxybutyl)azanediyl)bis(hexane- 6,1-diyl)bis(2 -hexyldecanoate)
•ALC- 0159: 2-[(polyethylene glycol) -2000] -N,N-ditetradecylacetamide
•DSPC: 1,2 -distearoyl -sn-glycero-3- phosphocholine
•Cholesterol
Diluent vials of sterile 0.9% Sodium Chloride Injection, USP are provided but shipped
separately by the applicant. The provided diluent is supplied as either a 10 mL or a 2
mL single- use vial. Alternate brand of sterile 0.9% sodium Chloride Injection, USP may
be used as the diluent and also for single use. The diluent vials should be discarded
after 1.8 mL is withdrawn.
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5.2 Manufacturers
Facilities and manufacturing sites involved in the commercial BNT162b2 DP
manufacturing and testing are presented in Table 23 below.
Table 23. Sites and Responsibilities for BNT162b2 Drug Product Manufacture
Site FEI/DUNS Numbers Responsibility
Pharmacia & Upjohn Company LLCc
7000 Portage Road
Kalamazoo, MI 49001
United States FEI: 1810189
DUNS: 618054084 LNP production/bulk DP formulation
Fill and finish
Primary packaging
Secondary packaging
Release and stability testing
Pfizer Manufacturing Belgium NV
Rijksweg 12
Puurs, 2870
Belgium FEI: 1000654629
DUNS: 370156507 LNP production/bulk DP formulation
Fill and finish
Primary packaging
Secondary packaging
Release and stability testing
Wyeth BioPharma Division of Wyeth
Pharmaceuticals LLCa
1 Burtt road
Andover, MA 01810
United States FEI: 1222181
DUNS: 17430868 Release and stability testing
Pfizer Inc.
875 Chesterfield Parkway West
Chesterfield, MO 63017
United States FEI: 1940118
DUNS: 004954111 Release and stability testing
Pfizer Ireland Pharmaceuticals
Grange Castle
Grange Castle Business Park
Clondalkin, Dublin 22
Ireland FEI: 30041455 94
DUNS: 985586408 Release and stability testing
FEI:
DUNS: Release testing (Sterility)
FEI:
DUNS: Release testing (Sterility)
a.The legal entity name change from Wyeth BioPharma Division of Wyeth Pharmaceuticals was
changed at the acquisition by Pfizer in 2009, since then the Wyeth Pharmaceuticals manufacturi ng
s
ite in Andover, Massachusetts belongs to Pfizer’s production sites and is embedded in Pfizer ’s
G
MP system. Pfizer will be utilized throughout the CTD
b. is a wholly owned subsidiary of Pfizer Inc.
c.Pharmacia & Upjohn Company LLC is a wholly owned subsidiary of Pfizer Inc.
Reviewer’s Comments:
Four manufacturing nodes we re employed for BNT162b2 DP manufacture at the time
of the initial EUA request: (1) Pfizer Puurs, (2) Pfizer Kalamazoo, (3) for
LNP formation/bulk DP formulation followed by fill and finish at Pfizer Puurs, and (4)
for LNP formation/bulk DP formulation followed by fill and finish
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at Pfizer Puurs. Additional DP manufacturing sites were introduced for emergency
supply production under EUA 27034, including Pfizer for DP fill and finish
and for LNP formation and bulk DP formulation.
For commercial BNT162b2 DP production, the Pfizer Puurs and Pfizer Kalamazoo
manufacturing facilities are included in the BLA submission. The Pfizer
site for the fi ll and finish will be added as a supplement after BLA approval.
Facilities and manufacturing sites for the production and testing/release of 0.9% sodium
chloride, USP diluent and their specified functions are listed in Table 24 below.
Table 24. Sites and Responsibilities for 0.9% Sodium Chloride, USP Manufacture
Site FEI/DUNS
Numbers Responsibility
Fresenius -Kabi USA, LLC
FEI#
DUNS# Manufacture, testing and
rel
ease (of 2 mL size diluent
vials)
Hospira, Inc
FEI#
DUNS# Manufacture, testing and
rel
ease (of 10 mL size
diluent vials)
Hospira is a wholly owned subsidiary of Pfizer Inc.
Reviewer’s Comments:
The saline diluent will be supplied as either a 10 mL single- use vial manufactured by
Hospira, Inc. (NDC 0409488810) or a 2 mL single- use vial manufactured by Fresenium
Kabi USA, LLC (NDC 6332318602). In amendment 47 submission (submitted on
August 13, 2021), the applicant listed an additional manufacturing facility, Pfizer
, for the production of the saline diluent. However, due to the
Warning Letter (reference WL issued on ) and O fficial Action
Indicated status, the agency will not approve the facility as the saline diluent supplier as
part of this BLA. The applicant acknowledged the request and removed Pfizer
. from the Diluent Manufacturers table in a follow -up
submission (amendment 56 submitted on August 17, 2021).
5.3 Control of Excipients
The compendial excipients used in the manufacture of BNT162b2 DP and their quality
standards are provided in Table 25 below.
Table 25 . Specifications for Compendial Excipients
Excipient Reference to Standard
Cholesterol USP-NF and/or Ph. Eur.
Sucrose USP-NF and Ph. Eur.
Sodium chloride USP-NF and Ph. Eur.
Potassium chloride USP-NF and/or Ph. Eur.
Dibasic sodium phosphate, dihydrate USP-NF and Ph. Eur.
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Excipient Reference to Standard
Monobasic potassium phosphate USP-NF and/or Ph. Eur.
Water for Injection USP-NF and Ph. Eur.
a. Specification also includes test for microbial contamination per USP<61> (Ph. Eur. 2.6.12)
Among the four lipid components used in the BNT162b2 LNP production, cholesterol is
a compendial excipient , and the other three lipids, ALC -0159, ALC -0315, and DSPC,
are non- compendial excipients. The supplier’s release specifications for all four lipids
include testing for
To ensure the quality of the lipids and
the resulting BNT162b2 DP, in- house control tests are performed for all four lipid
excipients including , prior to
release of the LNP DP. For cholesterol, an additional in- house test for
is also performed.
Table 26 below summarizes the source of the lipid excipients used for clinical study,
emer gency supply, and commercial BNT162b2 DP.
Table 26. List of Lipid Manufacturers with Phase of Use
Lipid Clinical Trial
Material Emergency Supply Commercial Supply
ALC-0315 Pfizer Groton,
, Pfizer Kalamazoo, Pfizer
Kalamazoo
ALC-0159 Pfizer Groton
DSPC
Cholesterol
Manufacturers’ full name s:
Pfizer, Groton, CT USA (Pfizer Groton);
Pharmacia & Upjohn Company LLC, Kalamazoo, MI USA (Pfizer Kalamazoo);
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5.4 Description of Drug Product Manufacturing Process and Process Controls
Flow diagrams for the DP manufacturing process and process controls are illustrated in
Appendices C and D for LNP production/bulk DP formulation and fill/finish operations,
respectively. For emergency supply/commercial DP manufacture, a scale- out approach
was used to increase capacity of the LNP - process with up to used
. The process scale approximates RNA per , such that
can be used to process as much as RNA correspondin g
to a bulk DP batch of approximately . The current batch size ranges at the two commercial DP manufacturing sites are set to be at Pfizer Kalamazoo and
at Pfizer Puurs.
The manufacturing process for the BNT162b2 DP includes the following major steps:
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Steps – Fill and Finish
For DP filli ng, sterile
bulk DP is aseptically filled into glass vials. At the end of filling, each vial is stoppered,
capped, and crimped. During filling, an in- process is performed at r outine
intervals for . All non- conforming vials are rejected.
Filled vials are 100% inspected for defects either through automated visual inspection or manual visual inspection. Vials passing inspection are statistically sampled for meeti ng
Acceptance Quality Limit (AQL) testing.
The labeling and packaging of the vaccine vials for commercial distribution is performed
on a fully automated or semi -automated packaging line. After labeling, the vials are
placed into trayboxes, and the boxes are manually closed and labeled.
Packaged BNT162b2 vials are frozen and stored in a freezer at - 90°C to - 60°C.
Critical- Process Parameters and In- Process Controls
To ensure DP manufacturing process consistency and the quality of the BNT162b2 DP,
relevant process parameters and IPT- Cs with acceptable ranges/acceptance criteria
have been established (Table 31). The critical process parameters (CPPs) were
conservativel y identified using Cause and Effect (C&E) Risk Assessment based on
product and process understanding, scientific rationale, and manufacturing experience and available data. IPT- Cs are used during the manufacturing process to control a
quality attribute/critical quality attribute within a specified range. In addition, IPT -Ms are
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implemented throughout the process to ensure a continuous state of control and to
enable forward processing.
Table 31. Process Control s for the Manufacture of BNT162b2 DP
Unit Operat ion Process Control/parameter Acceptance Range/
Acceptance Criteria Category
Kalamazoo Puurs
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Reviewer’s Comment:
Following further review of the parameters evaluated during process validation
at Pfizer Puurs, the following two process parameters were tightened as submitted in
BLA125742/0 amendment 19 on July 28, 2021:
The c
ontrol limits applied for testing and the process controls for
capping were reviewed by the facility reviewer.
Hold Times During DP Manufacturing Process
The hold times of the DP in- process materials during
are provided in Tables 32 and 33 ,
respectively.
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Ho
ld time for step was tightened from with up to
of this time allowed at up to with up to
of this time allowed up to . This change was made to ensure that the revised
-acceptance criterion for DP can be met. The process hold time change
has been validated via cumulative hold- time PPQ execution.
Hold Time Out of Freezing for the BNT162b2 DP
The Table 34 below summarizes the allowable times out of intended storage condition
of -90 to - 60°C during manufacture, packaging and transport.
Table 34. Time Out of Storage Condition for BNT162b2 Drug Product
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Hold Time for the Diluted BNT162b2 Vaccine Prior to Administration
The BNT162b2 final vaccine product is a concentrated suspension prepared in a
multidose vial free of preservatives. Following dilution with saline, the in- use hold period
and storage condition for the vaccine product is set to be up to 6 hours at 2°C to 25°C .
To support this hold condition, an in- vial dilution hold- time study was conducted in which
the physicochemical stability of the BNT162b2 DP, held in glass vials and diluted with saline as intended, was evaluated at 2-8°C, 25°C, and C. The results indicate that
the diluted vaccine can be stored at ambient temperature (25°C) for hours and can be exposed to elevated ambient temperature ( ) for hours with no impact on
product quality. In addition, a microbial challenge with selected model diluents and
surrogate DP solutions as well as a BNT162b2- specific microbial challenge were
conducted to evaluate the potential for microbial growth in a matrix representative of saline -diluted vaccine. In both studies,
microorganisms listed in and growth of
the microorganisms was monitored. The results demonstrate that at , which is
the proposed in- use time, no increase in growth was observed for any of the
organisms with the spiked test samples stored at 20- 25°C.
Reviewer’s Comments:
The result was close to the acceptance criterion of increase from T
0 for
at the -hour time point ( ) . Overall,
the results support the proposed in- use period of 6 hours at ambient temperature
after dilution with saline.
5.5 Process Validation
A global approach to development has been undertaken across multiple manufacturing
facilities in order to maximize BNT162b2 vaccine production and availability. The
process -validation approach for all DP supply nodes included within the
emergency/commercial supply network is composed of two phases. The Phase I
validation covers the overall DP manufacturing network by performing minimally one (1)
process validation (PV) run of each manufacturing supply node. The Phase II validation
covers the full validation of each of the supply nodes with separate protocols at each
site. Note that for the BLA submission, only Pfizer Puurs and Pfizer Kalamazoo are the proposed DP manufacturing sites for licensure.
Phase I Network PPQ Validation
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Hold Time for Packing, Shipping and Point of Use
DP Shipping Conditions of - 90 to - 15°C with an Allowance for a Maximum Cumulative
Time of at
The shipping condition of - 90 to -15°C with an allowance for a maximum cumulative
time of at is supported by the following:
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5.6 DP Manufacturing Process Development
Introduction of a Scaled- up Process for the Production of LNPs
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5.
7 Impurity Profile of the Drug Product
Possible process -related impurities include , introduced from
the DP manufacturing process, and , which are part of the ,
may be present in the final vaccine container. All these impurities are present in low
amounts and are further reduced during the DP manufacturing process by
.
Part of the process validation included evaluation of consistent removal of process -
related impurities throughout the manufacturing process. Results from the BNT162b2
DP process validation lots have demonstrated robust and consistent removal of
for all PPQ lots made at Pfizer Puurs and Pfizer Kalamazoo,
which is well below the safety concern limit of
. For , a comprehensive safety risk assessment has
been performed to compare the theoretical worst -case concentration of impurities that
could be introduced into the final DP assuming no impurity clearance during the manufacturing process. The evaluation results as shown in Table 4 2 below demonstrate
that none of the impurities pose a safety concern. Additionally, removal of was further demonstrated during process validation (quantification limit of in all PPQ BNT162b2 DP lots, which is equivalent to ).
Overall, based on t
he demonstration of consistent removal and the safety -based risk
assessment, all the process -related impurities from the DP manufacturing process do
not necessitate testing as part of BNT162b2 DP release.
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5.8 Characterization Studies on Drug Product
The DP characterization assays were developed to further describe and demonstrate
the structure and physicochemical properties of the DP. The enhanced analytical
characterization assays employed for the BNT162b2 DP and the testing results are briefly described as follows:
5.9 Control of Drug Product
Specifications
The specifications for BNT162b2 DP at release and throughput shelf life are shown in
Table 43 below.
Table 43. BNT162b2 Drug Product Specifications
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Quality Attribute Analytical Procedurea Acceptance Criteria
Appearance Appearance (Visual) White to off -white suspension
Appearance (Visible
Particulates) Appearance (Particles)
May contain white to off -white
opaque, amorphous particles
LNP
LNP
RNA
RNA Content
ALC-0315 Content
ALC-0159 Content
DSPC Content
Cholesterol Content
Vial Content (Volume)
Lipid Identities
Identity of Encoded RNA
Sequence
RNA
Bacterial Endotoxin Endotoxin
Sterility Sterilitye
No growth detected
Container -Closure Integrity Pass
a. All assays performed on stability unless otherwise noted
b. In accordance with , with minor difference in instrument calibration
c. Assay not performed on stability
d. Acceptance criteria values reflect correction
e. sterility test, which is performed in accordance with the with the exception of
method, may also be used
f. Tested at release and on stability for stability batches only
The acceptance criteria used for stability during shelf life will be predominantly the same
as the acceptance criteria used for lot release, with the exception of the LNP and RNA attributes, for both of which a separate stability acceptance criterion has
been established to enable alternative storage at - 20°C and 2- 8°C at the point of
administration.
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Reviewer’s Comment:
•The proposed commercial acceptance criteria for the following attributes at release
and during stability have been tightened for the commercial BNT162b2 DPcompared with the DP product under the EUA. The changes were made based onthe capability of the manufacturing process, historical release data for theemergency supply/commercial lots, as well as considerations of the assay
variability.
Justification of Specifications
The acceptance criteria in the DP specification reflect the current understanding of
criticality of quality attributes, their impact on product performance, and the quality of the product used in clinical trials to ensure consistent manufacture of DP. The lots used in the establishment of the commercial specification include nonclinical toxicology lot (
), clinical lots manufactured from process 1 DS ( ), and commercial -scale
emergency supply DP lots manufactured from process 2 DS ( lots manufactured between August 2020 and January 2021, including PPQ lots with manufactured at the Pfizer, Kalamazoo facility and manufactured at the Pfizer, Puurs site). Statistical analysis was applied to the release data for the commercial -scale DP lots. The
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mean, standard deviation and mean ± k*SD (k is the factor) for the release data set
were calculated for each quality attribute, if applicable. Based on the statistical analysis
results, the acceptance criteria were further adjusted and justified. For those attributes
not subjected to statistical analysis, acceptance criteria were determined based on understanding of formulation robustness, compendial requirements, clinical experience and/or literature references.
Table 44. Justification of Specifications for BNT162b2 Drug Product
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5.10
Analytical Procedures for Drug Product
The testing sites that are involved in assay performance and validation activities are
listed in Table 46 below.
Table 46. BNT162b2 Commercial D P Testing Sites
Analytical Procedure Testing Site Release, Stability
Appearance ( Visual )
Appearance ( Particles ) ARD , PGS -Puurs, PGS -KZO Release, Stability
ARD, PGS -GC, PGS -KZO Release, Stability
ARD , PGS -Puurs, PGS -KZO Release, Stability
ARD , PGS -Puurs, PGS -KZO Release
ARD , PGS -Puurs, PGS -KZO Release, Stability
ARD , PGS -Puurs, PGS -KZO Release, Stability
ARD , PGS -Puurs, PGS -KZO Release, Stability
Container Content ARD , PGS -Puurs, PGS -KZO Release
ARD, PGS -AND , PGS -GC Release
ARD, PGS -AND , PGS -GC Release, Stability
ARD , PGS -Puurs, PGS -KZO Release, Stability
Endotoxin PGS -Puurs, PGS -KZO Release, Stability
Sterility PGS -Puurs, PGS -KZO Release, Stability
Sterility PGS -Puurs, PGS -KZO Release, Stability
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• Pfizer Analytical Research and Development (ARD) Laboratories at Chesterfield, MO
(ARD-STL) and at Andover, MA (ARD -AND)
• Pfizer Global Supply, Andover, MA (PGS-AND)
• Pfizer Global Supply, Grange Castle, Ireland (PGS -GC)
• Pfizer Global Supply, Kalamazoo, MI (PGS- KZO)
• Pfizer Global Supply, Puurs, Belgium (PGS-Puurs)
Appropriate analytical procedures were established to monitor and assess the
BNT162b2 DP as detailed below.
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5
.11 Validation of Analytical Procedures
Compendial procedures were qualified for use in accordance with the applicable
pharmacopeias, excluding endotoxin and sterility. Endotoxin, in alignment with
, and sterility in alignment with
, were validated in the course of release testing of the CTM batches. The
validation of bacterial endotoxins and sterility tests are reviewed by DBSQC reviewers.
All the analytical procedures used for the BNT162b2 DP have been validated. The
assay may b e originally validated or co- validated in selected testing sites. Upon transfer
to additional testing location, reproducibility is evaluated in the validation or transfer.
A summary of the parameters evaluated for each analytical assay, the acceptance
criteria and the results are described in the following tables.
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.13 Summary of Batch Analysis
The BNT162b2 DP lots included in the section were used for nonclinical toxicology,
clinical trials, process performance qualification (PPQ), emergency supply, and stability.
The batch analysis data are presented for the DP lots manufactured across multiple
manufacturi ng sites, including from sites that are not being licensed under the BLA
submission . All batches met the specification at the time of release. Overall, the results demonstrate the capabilities of the manufacturing process for consistent production of quality DP.
The PPQ batch range data at Pfizer Puurs and at Pfizer Kalamazoo are shown in the Table 38.
5.14 Container -Closure System of the Final Container
The primary container -closure system for the BNT162b2 vaccine consists of the vial
components listed in Table 47.
Table 47. List of Components in Container Closure System
Component Description
Vial 2 mL Type I borosilicate glass vial, 13 mm finish
Vial Stopper 13 mm vial stopper composed of gray elastomer
(bromobutyl rubber) coated with a
Vial Seal 13 mm aluminum vial seal with tamper -evident polypropylene flip off cap
a. lubricant complies with requirements for
requirements for . requirements for Used as a Lubricant,
Extractables for the DP Container -Closure System
Controlled extraction studies were performed on the product contact bromobutyl rubber
stopper material ( ) using model solvents, including
. In one extraction study,
stoppers were extracted
. In a second extraction study, stoppers were
. For both extraction studies, volatile,
semi -volatile, non- volatile and elemental extractables were analyzed.
A Safety -Concern Threshold (SCT) was initially defined as total daily intake
(TDI) for each compound, a level at which unidentified or identified leachable compound presents negligible safety concern to patients. The putative hazard of each potential leachable compound was further assessed and the SCT adjusted accordingly based on the presence or absence of a mutagenic concern. Potential leachable compounds
without a mutagenic concern are assigned a SCT of TDI. Based on the
results from the extraction studies, the following compounds and elements were selected and monitored as potential leachables:
.
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Leachables for the DP Container -Closure System
Leachable studies are in progress to support the BNT162b2 commercial container
closure system with representative DP lots
. Vials stored at - 90°C to - 60°C are being tested using methods
validated for the potential leachables (identified in the extractables studies as well as
unexpected compounds) at the initial timepoint, 6 months,
. Currently available leachables information, including detection limits for
validated methods and initial time point results from DP lots, demonstrated that all
leachable compounds are below the SCT of TDI. Among the tested potential leachables, only were detected at
, respectively. In addition, no unidentified leachable
compounds have been detected. Based on a comprehensive review of available safety data, the presence of elements at or below the method quantitation limits in BNT162b2 DP pose negligible risk to patients. The rest of the leachable data will be reviewed in the
next GMP inspection.
5.15 Stability of Drug Product
At Pfizer Kalamazoo, the date of manufacture for the final drug product is defined as the date of the final sterile filtration operation. At Pfizer Puurs, however, the date of
manufacture is defined as the date of
. In either case, the date
of manufacture is no later than the date of final sterile filtration.
The proposed initial commercial shelf life of the BNT162b2 DP is 9 months when stored
at the intended storage condition of - 90°C to -60°C. The initial shelf life is determined
based on up to 9 months of currently available stability data on emergency supply
lots, and up to months of stability data on clinical and non- clinical lot s. The stability
data generated to date on the emergency supply and PPQ lots also support an
additional storage condition at - 20 ± 5°C for up to 2 weeks, as well as short -term
storage at 5 ± 3°C for up to one month (within the month shelf life).
The final commercial shelf life of the BNT162b2 DP is intended to be months when stored at the intended storage condition of - 90°C to - 60°C. The applicant commits to
subm itting stability data to support extension of the DP shelf life beyond the 9- month
period at intervals ( months) when supporting data are available.
Materials used for the DP stability studies include emergency supply and PPQ lots manufactured by Pfizer Puurs ( ), Pfizer Kalamazoo ( ),
(with fill/finish at Pfizer Puurs, ), and (with fill/finish at Pfizer Puurs, ), and clinical and non- clinical DP manufactured by .
The stability protocols for emergency supply/PPQ lots are described in Table 48 for the
long- term -90°C to - 60°C and the accelerated - 20 ± 5°C and 5 ± 3°C conditions.
Table 48. Stability Protocol for BNT162b2 Drug Product
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-90 to -60°C
(long -term)a-20 ± 5°C
(Accelerated)e 5 ± 3°C
(Accelerated)g
Appearance (Visible) 0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
Appearance (Visible Particulates) 0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
(ALC -0315 Content) 0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
(ALC -0159 Content) 0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
(DSPC Content) 0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
(Cholesterol Content) 0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
(RNA ) 0, 1Wd, 2W, 1M, 2M,
3M, 6M, 9M, 0, 1W, 2W, 0, 1W, 2W,
1M,
0, 3Mc, 6M, 0, 0,
Container Closure Integrity Test 0, Not Tested Not Tested
Endotoxin 0, Not Tested Not Tested
Sterility 0, Not Tested Not Tested
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a. Testing not performed at the 1W, 2W, or 2M time point for lot s
; Testing not performed at the 2M point for lots
; Testing not performed at the 1W or 2W time point for lot
b. Being performed at 3M and 6M time points for lots
c. Being performed at 3M time point for lots
d. 1W testing performed on lots
e. 1W testing performed on lot only; Testing not performed at the 1W, 2W, or time point for
lots ; Testing
not performed at the 1W or 2W time point for lots
f. Testing not performed on lots
g. 1W testing performed on lots ; 1M,
testing performed on lots ;
testing not performed on lots ; 1M and testing
performed on lots ends at time point
h. Testing not performed on lots
W = Week, M = Month
For emergency supply/PPQ lots, the updated stability results include the following:
• Lots : up to 9 months of stability data at the intended storage
condition of - 60 to -90°C , up to of data at the accelerated condition of -20
± 5°C, and up to months of data at the accelerated condition of 5 ± 3°C. For Lot
, up to of data are available at the stress condition s of
and
• Lots : up to of
stability data at the intended storage condition and the accelerated conditions of - 20
± 5°C, up to of data at the accelerated condition of 5 ± 3°C, as well as up
to of data at the stress condition
• Lots : up to of stability data at
all three tested storage conditions ( -60 to -90°C, -20 ± 5°C and 5 ± 3°C)
• Lots : up to of stability data at all three
tested storage conditions ( -60 to -90°C, -20 ± 5°C and 5 ± 3°C), For lots , up
to of stability data are available at the stress conditions of
• Lots : up to of stability data at all three
tested storage conditions ( -60 to -90°C, -20 ± 5°C and 5 ± 3°C) .
For clinical and non- clinical DP lots, the updated stability results include the following:
• Lots EE3818, ED3938, BCV40720- C, BCV40720- A, BCV40620 -E, and BCV40620-
A: up to 6 months of stability data at - 70 ± 10°C and up to of stability data
at 5 ± 3°C
• Lot BCV40420- A: up to months of stability data at -70 ± 10°C and up to
of stability data at 5 ± 3°C
• Non-clinical lot : up to of stability data at - 70 ± 10°C and
5 ± 3°C .
Summary of Stability Data
Long- term storage stability
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(b) (4)
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(b)
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105
For emergency supply and PPQ lots stored at - 90°C to -60°C and for clinical and non-
clinical lots stored at -70 ± 10°C, all results, generated to date, met the acceptance
criteria at the time of testing. Stability data from up to
months of data available) and emergency lots ( , up to 9
months of data available) support the proposed shelf -life of 9 months at the intended
storage conditions of - 90°C to - 60°C . Additionally, trending on critical stability -indicating
quality attributes (LNP , LNP , RNA content, RNA , lipid
content, RNA ) has been analyzed for stability lots and the data show
that there are no significant trends that would impact the shelf life of the DP through the
9-month time point when stored at the long- term condition of -90°C to - 60°C .
Accelerated stability
Accelerated stability
Accelerated -20 ± 5°C stability
All PPQ and eight emergency supply lots have been placed on formal stability at the accelerated - 20 ± 5°C storage condition. All data generated to date were within
the stated specifications through at least the time point. Out of the specification
results were were observed for LNP from multiple PPQ/emergency supply lots beginning from the time point.
Reviewer’s Comments:
Even though all stability data generated through the time point for DP lots
stored at -20 ± 5°C are within the specifications, trending for LNP is observed.
The available stability data showed that, except for , all
other lots (n = ) demonstrated an in LNP by . At the
time points, the trending is more evident, with the LNP ranging
from , which represents an average when
compared with the LNP at release that ranges from .
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
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(b) (4)
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(b) (4)
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106
Accelerated 5 ± 3°C stability
For emergency supply and PPQ lots, all data generated to date are within the
specifications through at least the 1- month time point , with the exception of
, both of which did not meet the specification of RNA (2- week time point
for and 1- month time point for ). Both lots were
manufactured using a lot of ALC -0315 made from the original lipid manufacturing
process. The process was later updated by
, and since the change, the resulting BNT162b2 DP all met the specification of
RNA when stored at 5 ± 3°C at all timepoints tested to date, which are 1 month for some lots and months for other lots.
For clinical DP lots, out -of-specification results were observed for RNA
beginning from the -month time point, and for LNP and LNP
beginning from the -month time point. It is not unexpected to see trends under the
accelerated storage condition and the results have no impact to the overall stability study.
Thermal Stress stability
Reviewer’s Comments:
As all the emergency supply BNT162b2 DP lots included in the stability protocol were
made using the commercial process, their stability data are being used to support the commercial use DP expiry of 9 months when stored at the intended storage condition
of -90 to - 60°C. A dditionally, the stability data generated to date support the DP shelf
life of up to two weeks at - 20 ± 5°C and up to 1 month (31 days) at 5 ± 3°C.
Additional Stability Studies for the BNT162b2 DP
stability
(b) (4)
(b) (4)
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(b) (4)
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107 s
tudies have been initiated for the emergency supply and PPQ lots to
provide support to the in- use period for the DP. The design of the thermal cycling
studies includes the following:
P
hotostability
was subjected to the photostability condition ( ).
DP vials were exposed to a light source providing an overall illumination of not less than
Post Approval Stability Protocol and Commitment
Upon completion of the stability protocol, post -approval, a minimum of of
BNT162b2 DP manufactured will be enrolled in the commercial stability program at the
long- term storage condition of - 90°C to - 60°C each year. The post -approval commercial
stability protocol for the BNT162b2 DP is described below in Table 49.
Table 49. Post -Approval Commercial Stability Pr otocol for DP Stored at - 90 to - 60°C
Analytical Procedure/Quality Attribute Test Intervals (Months)a
Appearance (Visible) 1, 6,
Appearance (Visible Particulates) 1, 6,
1, 6,
(b) (4)
(b) (4)
(b)(4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b)(4)
108 Analytical Procedure/Quality Attribute Test Intervals (Months)a
1, 6,
LNP 1, 6,
LNP 1, 6,
RNA 1, 6,
RNA Content 1, 6,
ALC-0315 Content 1, 6,
ALC-0159 Content 1, 6,
DSPC Content 1, 6,
Cholesterol Content 1, 6,
1, 6,
RNA 1, 6,
Container Closure Integrity Test Annually through end of shelf life
Sterility 0, End of shelf life
Endotoxin 0, End of shelf life
a.Additional test intervals may be included for the purpose of extending expiry.
6.Other CMC -related Information Not Covered in Module 3
Environmental Assessment or Claim of Categorical Exclusion
The applicant states that the pharmacologically active moiety or DS in COMIRNATY is
messenger RNA (mRNA), which is recognized as naturally occurring. Therapeutic use
of the vaccine product will not significantly alter the concentration or distribution of
mRNA, its metabolites, or degradation products in the environment.
The vaccine contains four pharmacologically inactive lipid excipients. The phospholipid
(DSPC) and the sterol lipid (cholesterol) are both recognized as naturally occurring membrane lipids, therefore, the use of the vaccine product will not alter significantly the
concentration or distribution of these lipids, their metabolites, or degradation products in
the environment. The other two novel lipids ALC -0315 and ALC -0159 are not
recognized as naturally occurring. However, the presence of both ALC -0315 and CLA -
0159 lipids is not expected to have an impact on the environment based on the conservatively estimated concentrations of each lipid at the point of entry into the aquatic environment (expected introduction concentration of for ALC -0315
and for ALC -0159).
Exception to the 21 CFR 610.15(a) Requirement for a Preservative
Under 21 CFR 610.15(a), a vaccine product in multiple- dose containers should contain
a preservative. On July 15, 2021, the applicant submitted to STN 125742/0 in amendment 11, a request for exception to the 21 CFR 610.15(a) requirement. The
justification for the unpreserved multi -dose presentation of the BNT162b2 vaccine
product includes the following:
• , the multi -dose preservative- free vial
presentation remain an important tool to enable sufficient global supply to deal with
the COVID -19 emergency.
(b)(4)
(b) (4)
(b) (4)
(b)(4)
(b) (4)
(b) (4)
(b)(4)
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(b) (4)
109 •The B
NT162b2 vaccine product is frozen at -90 to - 60°C for storage and distribution
with provisions for short -term storage for up to two weeks at - 20 ± 5°C and up to 1
month at 2 - 8°C until administration. On the day of administration, sterile saline
diluent is added into the thawed vial to provide 6 doses of the vaccine. Any unused
vaccine must be discarded 6 hours after dilution.
•The risks of this multi -dose preservati ve-free approach have been assessed by
taking into consideration of formulation factors, including
, DP storage temperature, and solution properties, which may impact the
ability of the finished DP to support or inhibit microbial growth. Dilution and
administration risks have also been evaluated from prior extensive experience with
other products and data from platform formulations and commonly used
studies. The microbial challenge assessment using the panel of m icrobes
described in support the in- use period of 6 hours to ensure adequate time
to prepare and administer 6 doses of the vaccine.
•Successful distribution of more than doses of vaccine in this multi-
dose, non- preserved presentation under the US EUA 27034 and other global
authorizations.
Reviewer’s Comments:
The request for an exception to the 21 CFR 610.15(a) for the BNT162b2 vaccine as a multi -dose preservative- free presentation is considered acceptable.
UNII code designations
The UNII code designations were reviewed and they appear to be acceptable.
7. Follow -up on Ongoing Issues Identified at the Time of EUA Request
RNA of BNT162b2 Drug Product –
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b)(4)
One page has been determined to be not releasable: (b)(4)
111 Intrinsic Particles During Visual Inspection
At the time of the EUA request, the sponsor reported that during the visual inspection
step of the DP manufacturing process, white- colored particles have been observed to a
varying degree across many lots spanning multiple manufacturing sites, including
and two fill/finish sites and across different sources (vendors
and batches). Upon investigation by , the
visible part icles were identified to be composed of
, and thus intrinsic to the product. Based on these observations, the
specification for appearance (visible particulates) was updated from “essentially free from visible particulates” for clinical lots to “may contain white to off -white opaque,
amorphous particles” for emergency supply/commercial lots.
Vials containing particles will be rejected and discarded during the 100% automated or manual inspection and Acceptable Quality Limit (AQL) sampling procedure for the inspected vials are further conducted to assess the robustness of the inspection method. The percentage of vials rejected due to particles during visual inspection has been low, ranging from . Additionally, visible particles are rarely observed
in vials during routine release or stability testing, indicating a low propensity for particle formation post -inspection. Further evaluation of samples from DP lot with and
without visible particles demonstrated that product quality (RNA content and
) was not impacted by the presence of visible particles.
Although visible particles may occasionally be observed in undiluted vials, following
dilution with sterile 0.9% sodium chloride and mixing, the dosing solutions are expected
to be free from visible particles. Any vials with visible particles observed after dilution should not be administered and should be discarded.
Reviewer’s Comments:
The impact of visible intrinsic particles appears to be minimal due to the following
reasons: (1) the particles are intrinsic to the product (i.e., they are not foreign
particles); (2) the frequency of occurrence is low; (3) the rare vials with intrinsic
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b)(4)
112 par
ticles can be identified during 100% visual inspection and discarded; (4) the vials
with intrinsic particles are not associated with changes in RNA content or
; (5) the intrinsic particles are dispersible upon dilution with saline; and
(6) given that the particulates are composed of the , the toxicity of which has
been characterized in non- clinical repeat -dose toxicity studies, and their presence as
visible particulates would not be associated with any unique chemical toxicological concern. In addition, as a precautionary measure, the instruction for the preparation of vaccine directs the healthcare provider to withhold administration if visible particles
are observed after dilution with saline.
8.Pre-Approval Inspection
The pre-a pproval inspection (PAI) for the Pfizer Puurs facility was conducted from June
24, 2021 through July 2, 2021. No 483 items were issued. The facility is considered
capable of consistently manufacturing the BNT162b2 DP with acceptable quality.
The PAI for the Pfizer Kalamazoo facility was waived. In May 2021, ORA/OBPO
performed a Level I surveillance inspection with focus on the production of the
BNT162b2 DP. A form 483 was issued; the observations were deemed Voluntary Action Indicated (VAI) based on the adequacy of the firm’s responses.
The PAI for the Pfizer Andover site was performed from July 19, 2021 through July 23,
2021. A form 483 with thirteen (13) observations was issued. On August 2, 2021, the
applicant submitted the response to STN 126742/0.25 to address all the 483 issues
identified during the Pfizer Andover PAI. The inspection was classified as VAI based on
the adequacy of the firm’s response to the Form 483.
During the PAI at Pfizer Andover, a recurring issue of out-of-specification
(OOS) in DS batches was noticed. An expedited investigation effort was made by the firm focusing on evaluating raw materials, operation parameters, and stability data. The
investigation was still ongoing and no definite root cause was yet identified during the inspection.
9.Nonclinical Studies
This review focuses on selected studies relevant to nonclinical pharmacology and
pharmacokinetics. Nonclinical toxicity studies and results were reviewed by the
toxicology reviewer.
9.1 Nonclinical Pharmacology Studies
Two variants of BNT162b2, “variant 8” and “variant 9” (V8 and V9, respectively) were
tested in nonclinical studies. The V8 and V9 variants differ only in their codon
optimization sequences but have the same amino acid sequence. Immunogenicity of BNT162b2 (V9) was evaluated in mice, rats, and nonhuman primates. Immunogenicity
(b) (4)
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113
of BNT162b2 (V8) was evaluated in rat s. Howe ver, only BNT162b2 (V9) has been
evaluated in the clinic and therefore is the subject of this marketing application.
Immunological assays used in nonclinical pharmacology studies include the following:
• S1 and receptor -binding domain( RBD) -binding IgG ELISA in mouse and rat
studies
• SARS- CoV-2 pseudovirus -based neutralization assay (pVNT) in mouse and rat
studies
• Direct Luminex -based immunoassay (dLIA) for S1- binding IgG in nonhuman
primate ( NHP ) studies
• Authentic SARS- CoV- 2 neutralization assay in NHP s tudies
• Interferon γ (IFNγ )-specific ELISpot in mouse and NHP studies
• Intracellular cytokine staining flow cytometry in mouse and NHP studies
Immunogenicity in Mice (Study Report: R -20-0085)
Four groups of eight female B ALB/c mice (at least 6 weeks of age) were immunized
once on day 0 with 0.2, 1, and 5 µg of BNT162b2 or with buffer (control group), by IM
injection. Blood was collected on d ays 7, 14, 21, and 28 after immunization, and
antibody immune responses were analyzed by ELISA and pVNT. On day 28, spleens
were collected for splenocyte isolation and analysis of T- cell responses using IFNγ
ELISpot assays. In addition, Luminex assays and intracellular cytokine staining (ICS)
were performed to assess cytokine responses.
The results demonstrated that BNT162b2 was highly immunogenic in mice with strong antigen- binding IgG and high- titer neutralizing antibody responses together with a Th1-
phenotype CD4+ response as well as an IFNγ+, IL- 2+, CD8+ T -cell response after a
single immunization. Total IgG ELISA showed that the vaccine induced a strong, dose-dependent IgG response that recognizes both S1 and RBD . First detection of IgG
antibodies was possible 7 days after immunization for all animals throughout the groups with an increase of total antibody amount until day 28. All mice developed functional
neutralizing antibodies starting at 14 days after immunization, and the titer increased
until the final study day. In addition, by profiling the IgG subtypes, a balanced IgG2a/IgG1 response was detected for the two higher doses (1 and 5 µg), while the low
dose induced a response with higher IgG1 than IgG2 levels (0.2 µg). Stimulation of
fresh splenocytes with an S protein- specific overlapping peptide pool demonstrated
robust CD 4+ and CD8+ T- cell IFNγ responses , and a Th1- dominant profile was
demonstrated in quantification of cytokines (IL- 2 and IFNγ) in the corresponding culture
supernatants.
Immunogenicity in Rats (Study Reports: 38166 and 20GR142)
Male and female Wistar Han r ats received three weekly IM doses of 100 µg of
BNT162b2 (V8) in study 38166 or 30 µg of BNT162b2 (V 9) in study 20GR142. For both
studies, s erum samples were collected on d ay 17, two days after the 3rd administration,
and on d ay 38, the end of the study. For study 38166, the sera were analyzed by ELISA
for IgG that bound S1 and RBD as well as for SARS -CoV- 2-S pseudovirus neutralizing
114 ant
ibodies. For study 20G142, sera were only analyzed for SARS -CoV-2 neutralization
activities. After immunization with BNT16 2b2 (V8), animals developed high titers of
antigen- specific antibodies as well as neutralization titers. BNT162b2 (V9) also elicited
SARS- CoV-2 neutralizing antibody responses in both male and female rats at the end of
the dosing and recovery phases of the study. SARS -CoV-2 neutralizing- antibody
responses were not observed in animals prior to vaccine administration or in saline-
administered control animals.
Immunogenicity and SARS -CoV-2 Challenge in Nonhuman Primates (Study Report:
VR-VTR- 10671)
BNT162b2 was assessed for immunogenicity and for protection against an infectious
SARS- CoV-2 challenge in rhesus macaques. Groups of 2-4- year-old male rhesus
macaques were immunized IM with 30 or 100 µg of BNT162b2 or saline control on days
0 and 21. In the study, a panel of 38 SARS -CoV- 2 human convalescent sera (HCS) was
included as a currently assessable benchmark to evaluate the quality of the humoral immune response to the vaccine. S1-binding IgG and SARS -CoV- 2 neutralizing
antibodies (determined by NT50 titers) were readily detectable as early as 14 days after a single immunization, and levels substantially increased further following the second
immunization. On day 28, seven days after dose 2, at the 30- µg dose level, the
neutralizing geometric mean titer (GMT) reached 8- fold the GMT of the 38 member
panel of HCS; at the 100 µg dose level, the neutralizing GMT was 18- fold the HCS
GMT. The HCS were drawn from SARS -CoV- 2 infected individuals 18 to 83 years of
age, at least 14 days after PCR -confirmed diagnosis and at a time when individuals
were asymptomatic. A decline of both S1- binding IgG levels and neutralizing titers was
observed at the latest measured time point ( day 56) but remained above the
neutralizing GMT and the S1- binding geometric mean concentration (GMC) of the HCS.
As seen following mouse immunization, strong S -specific Th1- dominant INFγ
+ but
minimal IL -4 T-cell responses were detected in all immunized rhesus macaques after
the the second 30 or 100 µg dose of the BNT162b2. By intracellular cytokine staining analysis, there was a dose- dependent increase in S -specific CD4
+ T-cell response wi th
a strong Th1- bias evidenced by a high frequency of INFγ+, IL-2+, or TNF -α+ cells.
Notably, CD8+ T-cell responses were also detectable in BNT- 162b2- immunized
animals.
Groups of 2- 4 year old male rhesus macaques that had received two IM immunizations
with 100 µg BNT162b2 V9 (n=6) or saline (Control; n=3) 21 days apart were challenged
55 days after the second immunization with 1.05× 106 plaque- forming units of SARS -
CoV-2 (strain USA- WA1/2020), which were split equally between the intranasal (IN) and
intratracheal (IT) routes. The presence of SARS -CoV- 2 RNA was then measured by
RT-qPCR in bronchoalveolar lavage fluid, nasal swabs, and oropharyngeal (OP) swabs.
Viral RNA was detected in BAL fluid in all three control -immunized animals (2 on Day 3
and 1 on Day 6); however, no viral RNA was detected in BNT162b2- immunized and
SARS- CoV-2-challenged animals. For nasal and OP swabs, viral RNA can be detected
in most of the control -immunized animals at all tested time points (Day s 1, 3, and 6),
115 w
hereas it was only detected in BNT162b2- immunized macaques on Day 1 after
challenge and became undetected on Day 3 and onward. None of the challenged
animals showed clinical signs of significant illness, indicating that the 2-4- year-old male
rhesus challenge model is primarily an infection model for SARS -CoV-2 and not a
COVID- 19 disease model. No radiographic evidence of vaccine- elicited enhanced
disease was observed.
9.2 Nonclinical Pharmacokinetics (PK) Evaluation
Assessment included evaluating the PK and metabolism of two novel lipid excipients
(ALC- 0315 and ALC -0159) in the LNP and potential biodistribution of BNT162b2.
The biodistribution of BNT162b2 was evaluated using expression as a surrogate reporter in mice. expressing modRNA was formula ted like
BNT162b2 with the identical lipid composition to generate LNPs . Mice were injected
intramuscularly with a total dose of 2 µ g/animal of LNP, and expression was
measured in vivo following application at 6 h, 24 h, 48 h, 72 h, 6 d, and 9 d after
injection. expression was identified at the injection site at 6 hours after injection and was not detected after 9 days. Expression in the liver was also present to
a lesser extent at 6 hours after injection and was not detected by 48 hours after
injection. A tissue distribution study was also performed in Rats using
LNPs, which similarly comprised of a proprietary mixture of BNT162b2 lipid
components and -encoding mRNA. The test item also contains trace amounts
of radiolabeled , a non -ex
changeable, non- metabolizable lipid marker in order
to monitor the disposition of the LNP. The tested animals (21 male and 21 female) each
received a single intramuscular dose of at a target mRNA total dose of
50 µg/animal. Whole blood and tissue samples were collected at 0.25, 1, 2, 4, 8, 24 and
48 hours post -dose (three animals/sex/timepoint). The study results showed that
following intramuscular injection, the concentration of was gr eatest in
the injection site at all time point s. Outside the injection site, l evels of radioactivity
peaked in the plasma by 1- 4 hours post -dose and distributed mainly into liver, adrenal
glands, spleen and ovaries over 48 hours. Total recovery of radioactivity outside of the
injection site was greatest in the liver, with much lower total recovery in spleen and very little recovery in adrenals glands and ovaries. The mean concentrations and tissue
distribution pattern were broadly similar between the sexes.
An rat PK study, using the - encoding modRNA formulated in
an identical lipid composition as BNT162b2, was performed to evaluate
pharmacokinetics of both ALC -0315 and ALC -0159 lipids . In this study, plasma samples
were collected up to 336 hours following a single administration to male
Rats at a dose of 1 mg/kg. The results demonstrated that ALC -0315 and ALC -0159
distribute from the plasma to the liver. Plasma concentrations of ALC -0315 and ALC -
0159 decreased rapidly, with initial t
1/2 of 1.62 and 1.72 h, respectively. ALC -0315 and
ALC- 0159 were then cleared from plasma, resulting in terminal elimination t 1/2 of 139
and 72.7 h, respectively. The estimated percent of dose distributed to the liver was ~ 60% for ALC -0315 and ~20% for ALC -0159. While there was no detectable excretion of
(b) (4)
(b) (4)
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(b) (4)
(b) (4)
(b) (4)
(b) (4)
(b) (4)
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116 ei
ther lipid in the urine, the percent of lipids excreted unchanged in feces was ~1% for
ALC- 0315 and ~50% for ALC -0159.
The in vitro metabolism of ALC- 0315 and ALC -0159 was evaluated in blood, liver
microsomes, S9 fractions, and hepatocytes from mice, rats, monkeys, and humans. The
in vivo metabolism was examined in rat plasma, urine, feces, and liver samples from the
PK study. Metabolism of ALC -0315 and ALC -0159 appears to occur slowly in vitro and
in vivo . ALC- 0315 and ALC -0159 are metabolized by hydrolytic metabolism of the ester
and amide functionalities, respectively, and this hydrolytic metabolism is observed
across the species evaluated.
Reviewer’s Comments
Based on current hypotheses regarding the etiology of vaccine- associated enhanced
disease, the provided data are reassuring due to: (1) the robust induction of
functional (i.e., neutralizing) antibodies in mice, rats and rhesus macaques ; (2) the
Th1-bias in T -cell responses; and (3) the lack of disease in vaccinated rhesus
macaques challenged with SARS -CoV- 2. The nonclinical ADME studies indicate that
the LNP mainly localizes to the site of injection and, to a lesser extent, distributes to the liver. Approximately 50% of ALC -0159 is excreted unchanged in feces, while
metabolism appears to play a role in the elimination of ALC -0315.
10. Clinical Assays
10.1 Diagnostic Assays Used to Support Clinical Efficacy Endpoints
Two diagnostic assays (Cepheid Xpert Xpress RT- PCR assay for the detection of
SARS- CoV-2 in clinical specimens and Roche Elecsys Anti -SARS- CoV- 2 assay for the
evaluation of serostatus to SARS -CoV- 2) are described below.
Cepheid Xpert Xpress RT- PCR Assay
The Cepheid Xper t Xpress SARS -CoV- 2 assay, which has received FDA authorization
under an EUA, is a rapid, automated in vitro diagnostic test for the qualitative detection
of the nucleocapsid (N) and envelope ( E) gene sequences from nasopharyngeal, nasal,
or mid -turbinate swab and/or nasal wash/ aspirate specimens collected from patients
suspected of having COVID -19 disease. The Cepheid Xpert assay was used to assess
viral infection before vaccination and to confirm COVID -19 disease cases during study
follow- up. Detection of RNA sequences for the N and E genes are carried out by real -
time RT -PCR following a single -step sample processing protocol.
Report VR-MVR- 10080 describes the method validation for the Cepheid Xpert Xpress
PCR Assay conducted at Pfizer Pearl River . The validation results are summarized
below:
Detection limits : Based on simulated samples (generated by spiking with the
commercially available AccuPlex SARS- CoV-2 reference material or a live SARS- CoV- 2
reporter virus) , the detection limits was established to be and
.
(b) (4)
(b) (4)
(b) (4)
117 Accuracy: The assay’s clinical accuracy was evaluated using simulated samples,
patient samples as well as pre- COVID -19 samples.
Tak
en together, the validation results for the Cepheid Xpert Xpress SARS -CoV- 2 assay
support its intended use to test samples from Pfizer’ s clinical efficacy trial of the SARS-
CoV- 2 vaccine candidate and other epidemiological studies.
Roche Elecsys Anti -SARS- CoV- 2 Assay
The Roche Elecsys Anti -SARS- CoV- 2 assay, which has received FDA authorization
under an EUA, is a rapid, automated in vitro diagnostic test for detecting the presence
of antibodies to nucleocapsid (N) protein of SARS- CoV-2 (antigen not present in the
BNT162 b2 vaccine candidate) in serum or plasma samples. This is a qualitative assay
marketed as an aid in identifying individuals with an adaptive immune response to
SARS- CoV-2, which would indicate a recent or prior infection. The Elecsys Anti -SARS-
CoV- 2 assay will be used to assess serostatus before vaccination. Detection of N
protein antibodies to SARS -CoV- 2 is carried out by a two-step immunoassay method in
which biotin - and ruthenium -conjugates of N protein are incubated with human serum or
plasma. Streptavi din-coated paramagnetic beads are then added to isolate N
protein/antibody complexes and chemiluminescent emissions are measured by a
photomultiplier. Results are presented in qualitative “Positive” or “Negative” based on N -
specific antibody levels above or below a predetermined cutoff index (COI) value.
The specificity of the assay was evaluated using
(b)(4)
(b)(4)
118 The sensitivity of the Elecsys Anti -SARS- CoV-2 assay was evaluated
R
eview er’s Comments:
The submitted data are supportive of both the Cepheid Xpert Xpress assay and the
Roche Elecsys Anti -SARS- CoV- 2 assay being suitable for their intended use in
Phase 2/3 clinical studies to evaluate the efficacy of the BNT162b2 vaccine
perform ed at Pfizer’s testing facility (Pfizer Vaccine Research and Development;
Pearl River, NY). It is important to note that the validation studies relied on available
information generated by the assay kit manufacturers (Cepheid and Roche) in support of assay EUA as well as academic studies assessing assay performance
published in peer -reviewed publications; the validation results generated by the
applicant were in general agreement with information from these external sources.
10.2 Immunogenicity Assays Used for Exploratory Immunogenicity Endpoints
Two immunogenicity assays, SARS -CoV- 2 mNeonGreen virus microneutralization
assay and direct Luminex a ssay (dLIA) for IgG quantification, are used for
evaluating the immune responses from clinical trial samples.
SARS- CoV-2 mNeonGreen Virus Microneutralization Assay (SARS- CoV- 2 mNG NT)
The SARS -CoV- 2 mNG NT is a biofunctional assay that measures neutralizing
antibodies against SARS -CoV- 2. The SARS -CoV- 2 mNG virus is derived from the
USA_WA1/2020 strain that had been engineered to contain an mNeonGreen (mNG)
reporter gene in open reading frame 7 of the viral genome. The recombinant virus is
rescued by reverse genetics and upon productive infection of cells , green fluorescence
is produced. This reporter virus generates similar plaque morphologies and indistinguishable growth curves from wild- type virus.
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119 R
eport VR-MVR- 10083 describes the method validation for the SARS- CoV- 2 mNG NT
assay conducted at Pfizer Hackensack. The validation results are summarized below:
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eviewer’s Comments
Assa y specificity was assessed in the evaluation of the limit of detection. This
validation study at Pfizer Hackensack involved assessing
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120 D irect Luminex Assay (dLIA)
The S 1 IgG dLIA measures IgG antibody levels to the subunit 1 (S1) of the
SARS- CoV-2 spike protein in human serum samples.
R
eport VR -MQR- 10211 summarizes qualification results for the S1 IgG
dLIA. The qualification results are also summarized below:
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121
Reviewer’s Comments
The qualification study reports for the S1 IgG dLIA and RBD IgG dLIA w ere submitted
to the original BLA. Upon request for the validation reports for both assays, the
applicant submitted to STN125742/0 amendment 19, the dLIA validation report (VR -
MVR- 10077) for quanti fication of IgG antibodies to SARS -CoV- 2 full -length S protein.
Additional IR was issued on August 2, 2021 to obtain clarification on which
immunogenicity assay was used in the clinical studies. The applicant provided the response to amendment 31 on August 5, 2021, stating that t he S1 IgG dLIA was
used for immunogenicity assessments in Phase I (C4591001 sentinel cohort) and Phase 2 (C4591001 Phase 2 immunogenicity cohort) studies. The validated full -
length S -binding IgG dLIA will be used for the lot consistency study and is not part of
the original BLA filing.
The qualification study report for the S1 IgG dLIA assay is considered acceptable for
this BLA submission with the reasons as follows: 1) the vaccine efficacy is
determined by the clinical endpoints and the dLIA assay was used for the exploratory
immunogenicit y endpoint; and 2) appropriate assay parameters were evaluated
during the qualification study, including precision, linearity, lower limit of
detection/quantification, range, reference standard bias, and assay run performance.
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122
The qualification results for the S1 IgG dLIA assay support its intended use for the
quantification of anti -S1 IgG antibodies in human sera.
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