Document text
Principal Investigator: Julie Champion
Organization: GEORGIA INSTITUTE OF TECHNOLOGY
Fiscal Year: 2021
Award: $397,382
Funding agency: National Institute of Biomedical Imaging and Bioengineering
Abstract
SARS-CoV-2 has spread rapidly across the globe. While vaccines are in development and
existing drugs have shown some efficacy in reducing disease severity, there is an urgent need
for molecules that can serve in both a prophylactic and therapeutic function. Proteins are well
suited for this task because they can be engineered to have high-specificity and high-affinity for
targets, and genetic protein fusion bestows multiple functions on a single protein molecule. A
nanobody (Nb) has been identified with high affinity for the receptor binding domain (RBD) of the
SARS-CoV-2 spike protein, which mediates viral entry into cells via the angiotensin converting
enzyme 2 (ACE2) receptor. Nbs are small heavy chain, single domain camelid antibodies that are
stable over wide conditions, express well in multiple systems, can be humanized to reduce
immunogenicity, and can be fused to other proteins. The goal of this proposal is to develop a
protein-based therapeutic platform that can be taken either prophylactically or post-infection to
reduce the severity of viral infection. The overall hypothesis is that a self-assembling hexameric
coiled-coli (Hex) can be fused to 12-24 anti-RBD Nbs to create a complex with high avidity that
will neutralize virus and prevent entry and/or aggregate virus into traps that can be cleared.
Further, anti-albumin Nb will be incorporated in the fusion design to investigate any benefit of
increased circulation time. Three aims have been set to meet the goal and test the hypothesis.
(1) Design, fabricate and characterize Hex-Nb assemblies made from fusion proteins with
different numbers of anti-RBD and anti-albumin Nbs. (2) Characterize binding, neutralization, and
trapping of SARS-CoV-2 pseudovirus by Hex-Nb fusion assemblies in vitro. (3) Determine
pharmacokinetics, biodistribution and humoral immunogenicity of Hex-nanobody fusion protein in
mice following intravenous injection. From this work, an innovative protein-based therapeutic will
be constructed from functional nanobodies that inhibits SARS-CoV-2 pseudovirus infection and
extends in vivo lifetime, providing proof of concept for Hex assemblies to serve as prophylactics
or therapeutics to prevent or treat viral infections. These results will enable future preclinical work,
including viral challenge studies and direct airway administration of the Hex-Nb assemblies.
Overall, this work will provide a strategy to respond to both the current and future pandemics with
highly multivalent engineered protein complexes.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><ACE2><Affinity><Albumins><Amino Acid Sequence><Antibodies><Avidity><Bacterial Infections><Binding><Binding Sites><Biodistribution><Blood Circulation><Blood Circulation Time><Bloodstream><COVID-19 virus><COVID19 virus><Cancers><Cardiovascular><Cardiovascular Body System><Cardiovascular Organ System><Cardiovascular system><Cell Body><Cells><Cessation of life><Charge><Chimera Protein><Chimeric Proteins><Circulation><Clinical Treatment Moab><CoV-2><CoV2><Combining Site><Complex><Death><Development><Disease><Disorder><Drug Kinetics><Drugs><Engineering><Evaluation><Fusion Protein><Future><GS-5734><Genetic><Goals><Health><Health Care Providers><Health Personnel><Healthcare Providers><Healthcare worker><Heart Vascular><Human><In Vitro><Infection><Infection prevention><Inflammatory><Ligands><Malignant Neoplasms><Malignant Tumor><Measurement><Measures><Mediating><Medication><Mice><Mice Mammals><Modern Man><Molecular Interaction><Monoclonal Antibodies><Murine><Mus><Nature><Organ failure><Outcome><Patient outcome><Patient-Centered Outcomes><Patient-Focused Outcomes><Patients><Pharmaceutic Preparations><Pharmaceutical Preparations><Pharmacokinetics><Prevent infection><Prevention><Primary Protein Structure><Property><Protein Engineering><Proteins><Rapid screening><Reactive Site><Receptor Protein><Recombinant Proteins><Recombinants><Research><SARS><SARS corona virus 2><SARS coronavirus disease><SARS-CoV disease><SARS-CoV-2><SARS-CoV2><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome CoV disease><Severe Acute Respiratory Syndrome coronavirus disease><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome related corona virus 2><Severities><Severity of illness><Specificity><Structure><System><Technology><Temperature><Testing><Therapeutic><Time><Translations><Vaccines><Variant><Variation><Veklury><Viral><Viral Diseases><Virus><Virus Diseases><Vulnerable Populations><Work><Wuhan coronavirus><angiotensin converting enzyme 2><angiotensin converting enzyme II><bacteria infection><bacterial disease><base><block viral entry><circulatory system><coronavirus disease 2019 virus><design><designing><developmental><disease severity><driving force><drug/agent><effective therapy><effective treatment><genetic protein engineering><hCoV19><health care personnel><health care worker><health provider><health workforce><healthcare personnel><immune clearance><immune elimination><immunogenicity><improved><in vivo><inhibit viral entry><innovate><innovation><innovative><intravenous injection><mAbs><malignancy><medical personnel><nCoV2><nano meter scale><nano meter sized><nano scale><nanobodies><nanobody><nanometer scale><nanometer sized><nanoscale><native protein drug><neoplasm/cancer><new drug treatments><new drugs><new therapeutics><new therapy><next generation therapeutics><novel drug treatments><novel drugs><novel therapeutics><novel therapy><pandemic><pandemic disease><pharmaceutical protein><pre-clinical><preclinical><prevent><preventing><prophylactic><protein complex><protein design><protein drug agent><protein sequence><protein structure><protein structures><proteins structure><receptor><receptor binding><receptor bound><remdesivir><response><scale up><sdAb><single domain antibodies><small molecule><therapeutic effectiveness><therapeutic protein><treatment provider><treatment strategy><viral entry blocker><viral entry inhibitor><viral infection><virus infection><virus-induced disease><vulnerable group>