019736 S70 M1 response ir 29jul2020 clin assay qual

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SARS -CoV- 2 mRNA Vaccine (BNT162, PF -07302048)
BB-IND 19736
M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 1SARS -CoV -2 mRNA VACCINE (BNT162, PF -07302048)
BB-IND 19736
Response to CBER Request for Information Received on 29 July 2020 –
Questions Regarding Clinical Assay Qualification
21August 2020
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FDA-CBER-2021-5683-1072455
SARS -CoV- 2 mRNA Vaccine (BNT162, PF -07302048)
BB-IND 19736
M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 2TABLE OF CONTENTS
LIST OF TABLES ................................ ................................ ................................ ..................... 4
LIST OF FIGURES ................................ ................................ ................................ ................... 4
1. INTRODUCTION ................................ ................................ ................................ ................. 5
2. REQUESTS ................................ ................................ ................................ ........................... 5
2.1. CBER Request 1 ................................ ................................ ................................ ........ 5
2.1.1. Sponsor’s Response................................ ................................ ...................... 5
2.2. CBER Request 2 ................................ ................................ ................................ ........ 6
2.2.1. Sponsor’s Response................................ ................................ ...................... 6
2.3. CBE R Request 3 ................................ ................................ ................................ ........ 6
2.3.1. Sponsor’s Response................................ ................................ ...................... 6
2.4. CBER Request 4 ................................ ................................ ................................ ........ 7
2.4.1. Sponsor’s Response................................ ................................ ...................... 7
2.5. CBER Request 5 ................................ ................................ ................................ ........ 7
2.5.1. CBER Re quest 5a ................................ ................................ ......................... 7
2.5.1.1. Sponsor’s Response................................ ................................ .....7
2.5.2. CBER Request 5b ................................ ................................ ......................... 8
2.5.2.1. Sponsor’s Response................................ ................................ .....8
2.5.3. CBER Request 5c ................................ ................................ ......................... 8
2.5.3.1. Sponsor’s Response ................................ ................................ .....8
2.6. CBER Request 6 ................................ ................................ ................................ ........ 8
2.6.1. Sponsor’s Response................................ ................................ ...................... 8
2.7. CBER Request 7 ................................ ................................ ................................ ........ 9
2.7.1. Sponsor’s Response................................ ................................ ...................... 9
2.8. CBE R Request 8 ................................ ................................ ................................ ......13
2.8.1. Sponsor’s Response................................ ................................ .................... 13
2.9. CBER Request 9 ................................ ................................ ................................ ......14
2.9.1. Sponsor’s Response................................ ................................ .................... 14
2.10. CBER Request 10 ................................ ................................ ................................ ..15
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BB-IND 19736
M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 32.10.1. Sponsor’s Response................................ ................................ .................. 15
2.11. CBER Request 11 ................................ ................................ ................................ ..15
2.11.1. CBER Request 11a ................................ ................................ ................... 15
2.11.1.1. Sponsor’s Response................................ ................................ .15
2.11.2. Request 11b ................................ ................................ .............................. 15
2.11.2.1. Sponsor’s Response ................................ ................................ .15
2.12. CBER Request 12 ................................ ................................ ................................ ..16
2.12.1. Sponsor’s Response................................ ................................ .................. 16
2.13. CBER Request 13 ................................ ................................ ................................ ..16
2.13.1. CBER Request 13a ................................ ................................ ................... 16
2.13.1.1 . Sponsor’s Response................................ ................................ .16
2.13.2. CBER Request 13b ................................ ................................ ................... 16
2.13.2.1. Sponsor’s Response................................ ................................ .16
2.14. CBER Request 14 ................................ ................................ ................................ ..17
2.14.1. Sponsor’s Response................................ ................................ .................. 17
2.15. CBER Request 15 ................................ ................................ ................................ ..17
2.15.1. Sponsor’s Response................................ ................................ .................. 17
2.16. CBER Request 16 ................................ ................................ ................................ ..17
2.16.1. Sponsor’s Response................................ ................................ .................. 17
2.17. CBER Request 17 ................................ ................................ ................................ ..18
2.17.1. Sponsor’s Response ................................ ................................ .................. 18
2.18. CBER Request 18 ................................ ................................ ................................ ..18
2.18.1. Sponsor’s Response................................ ................................ .................. 18
2.19. CBER Request 19 ................................ ................................ ................................ ..18
2.19.1. Sponsor’s Response................................ ................................ .................. 18
2.20. CBER Request 20 ................................ ................................ ................................ ..18
2.20.1. Sponsor’s Response................................ ................................ .................. 18
3. REFERENCES ................................ ................................ ................................ .................... 19
4. SUPPORTING DOCUME NTATION ................................ ................................ ................. 20
4.1. New, Appended, or Replaced Supporting Documentation ................................ .....20
4.2. Previously  Submitted Supporting Documentation ................................ .................. 20
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SARS -CoV- 2 mRNA Vaccine (BNT162, PF -07302048)
BB-IND 19736
M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 4LIST OF TABLES
Table 1. SARS -COV -2 IgG d LIA –Calculated  for S1 Sample SAN96935 ............. 11
Table 2. S1 Protein – Neat Estimates ................................ .............................. 12
Table 3. SARS -CoV -2 RBD IgG Luminex Assay ................................ ............................. 13
Table 4. SARS -CoV -2 S1 IgG Luminex Assay ................................ ................................ .14
LIST OF FIGURES
Figure 1.  Linearity  of Sample SAN96935 for SARS -COV -2 IgG dLIA –
S1................................ ................................ ................................ .......................... 10
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SARS -CoV- 2 mRNA Vaccine (BNT162, PF -07302048)
BB-IND 19736
M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 82.5.2. CBER Request 5b
Please clarify how the pre-pandemic samples were used to assess precision.
2.5.2.1. Sponsor’s Response
In the RBD and S1 IgG Luminex assay  qual ifications, Pfizer used  sera and 
pre-pandemic sera to assess the assay  precision, as these were the sera available at the 
time.  The pre -pandemic serum samples had measurable low concentrations to S1 and RBD 
IgG that were reproducible. Excluding these samples does not appreciabl y affect assay 
precision (overall assay  precision changes from  and  for S1 and 
RBD, respectivel y).  
2.5.3. CBER Request 5c
Please submit the precision data in analyzable format, eg, excel, xpt , etc.
2.5.3.1. Sponsor’s Response
Previously  wesubmitted draft versions of the RBD and S1 IgG Luminex assay  
Qualifications.  The final versions of both the RBD and S1 IgG Luminex assay  
Qualifications are being provided in this submission.  Within the final qualif ication report, 
attachment 1 is included.  This attachment provides data tables that are inclusive of all assay  
results (including precision anal yses) obtained during the qualifications.  The precision data 
in an excel file are also included as requested.
2.6.CBER Request 6
Your qualification assessments did not include assessments of the performance of the 
assay at the lower range (ie, sensitivity) or determination of the lower limit of detection 
(LOD).  Please explain how the assay will be used (eg, if it wil l be used to categorize 
seropositive and seronegative samples).  If needed, establish an LOD based on the lowest 
concentration that may be detected above the background with a defined probability.
2.6.1. Sponsor’s Response
The RBD and S1 IgG Luminex assays are quantitative assay s used for measuring specific 
IgG levels in serum samples from subjects who have received a candidate SARS -CoV -2 
vaccine.  Limits of Quantitation (L OQ) are assessed.  Pfizer does not intend to use the RBD 
and S1 IgG Luminex assay s to categorize seropositive and seronegative samples as we plan 
to use a commercial test (Roche Elecsy s Anti- SARS CoV -2 submitted on 31 July  2020 under 
EUA2000514/A001) for that purpose; therefore, each assay ’s Limit of Detection (LOD) was 
not assessed.
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SARS -CoV- 2 mRNA Vaccine (BNT162, PF -07302048)
BB-IND 19736
M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 10Figure 1.  Linearity of Sample SAN96935 for SARS-COV- 2 IgG dLIA – S1
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BB-IND 19736
M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 11For S1 sample SAN96935 in the SARS -COV -2 IgG dLIA plotted in Figure 1,  
Table 1.SARS -COV-2 IgG dLIA – Calculated  for S1 Sample SAN96935
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BB-IND 19736
M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 12Table 1.SARS -COV-2 IgG dLIA – Calculated  for S1 Sample SAN96935
Table 2.S1 Protein – SAN96935 Neat Estimate s
Batch Analyst Plate Number Neat
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M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 172.14. CBER Request 14
In your validation protocol, please include details about the assay -critical reagents to be 
used in the validation studies, their source and preparation for use.  For each validation 
assessment, please describe the serum panels used and the dilution scheme including 
dilution matrix used.
2.14.1. Sponsor’s Response
Pfizer will include details about the critical reagents in the validation protocol.
2.15. CBER Request 15
We recommend that you submit the validation protocol for the selected dLIA(s) for our 
review before starting the validation studies.
2.15.1. Sponsor’s Response
Pfizer will submit the validation protocol prior to start of the validation studies. We like to 
note however, that since the SARS -CoV -2 Luminex assay s are platform assay s, we plan to 
validate these assay s in the same way  we have done successfully  in the past for other 
programs.  
REQUESTS 16-20: Regarding the qualification report for the SARS-CoV-2 mNeonGreen 
virus microneutralization (MN) assay (VR -MQR-10214)7performed at the University of 
Texas Medical Branch (UTMB):
Given that the qualification only included a preliminary precision assessment and trend 
analysis of control sera, at this time we are only providing general input for your 
consideration.
2.16. CBER Request 16
We acknowledge that in your current plans the MN assay will not be used to assess Phase 
3 endpoints to support licensure. However, since a neutralization assay is used to assess 
functional antibody, we strongly encourage that the assay be validated at your Hackensack 
Meridian Health site. 
2.16.1. Sponsor’s Response
Pfizer acknowledges CBER’s comment.  Pfizer intends to use the microneutralization assay  
to support exploratory  endpoints in the phase 2/3 stage of the C4591001 study  and will use 
the qualified assay  for this purpose.  Pfizer intends to validate the microneutralizatio n assay  
at the Hackensack Meridian Health site, if there is a potential need to support future licensing 
studies.
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SARS -CoV- 2 mRNA Vaccine (BNT162, PF -07302048)
BB-IND 19736
M 1.11.3 Response to FDA Information Request
PFIZER CONFIDENTIAL
Page 182.17. CBER Request 17
Please evaluate the linearity of your assay and provide the assay range for the test samples 
based on linearity and precision results.
2.17.1. Sponsor’s Response
Pfizer acknowledges CBER’s comment and will evaluate  linearity  of the assay  to 
inform the assay  range.
2.18. CBER Request 18
We are concerned with your precision assessment of the  the  is 
higher than your proposed target of  Please comment.
2.18.1. Sponsor’s Response
Pfizer acknowledges CBER’s comment. The relatively  high  for  
was impacted b y the low number of assay runs included in the dataset. The inclusion of 
additional datapoints from additional runs would be expected to lower the . Pfizer 
will re -evaluate QCS ranges prior to testing clinical samples at HMH.
2.19. CBER Request 19
For validation of the MN assay, please consider performing prelim inary qualifications at 
the laboratory where the assay will be validated and further developing the assay 
procedures and submitting the qualification information and validation protocol for our 
input.
2.19.1. Sponsor’s Response
Pfizer agrees that preliminary  asses sments of assay performance in the new laboratory  at 
Hackensack Meridian Health (where the assay  will be validated) is important and submit the 
requested data and the validation protocol for CBER’s review and comment.
2.20. CBER Request 20
We recommend that you submit the validation protocol for the SARS -CoV-2 MN assay for 
our review before starting the validation studies.
2.20.1. Sponsor’s Response
Pfizer will provide a validation protocol for CBER’s review and comment prior to validation.
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Page 204. SUPPORTING DOCUMENTA TION
None.
4.1. New, Appended, or Replaced Supporting Documentation
None.
4.2. Previously Submitted Supporting Documentation
None.
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