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Rapid identification of neutralizing antibodies against SARS-CoV-2 variants by mRNA display.

Cell reports (2022-02-04)
Shiho Tanaka, C Anders Olson, Christopher O Barnes, Wendy Higashide, Marcos Gonzalez, Justin Taft, Ashley Richardson, Marta Martin-Fernandez, Dusan Bogunovic, Priyanthi N P Gnanapragasam, Pamela J Bjorkman, Patricia Spilman, Kayvan Niazi, Shahrooz Rabizadeh, Patrick Soon-Shiong
ABSTRACT

The increasing prevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants with the ability to escape existing humoral protection conferred by previous infection and/or immunization necessitates the discovery of broadly reactive neutralizing antibodies (nAbs). Utilizing mRNA display, we identify a set of antibodies against SARS-CoV-2 spike (S) proteins and characterize the structures of nAbs that recognize epitopes in the S1 subunit of the S glycoprotein. These structural studies reveal distinct binding modes for several antibodies, including the targeting of rare cryptic epitopes in the receptor-binding domain (RBD) of S that interact with angiotensin-converting enzyme 2 (ACE2) to initiate infection, as well as the S1 subdomain 1. Further, we engineer a potent ACE2-blocking nAb to sustain binding to S RBD with the E484K and L452R substitutions found in multiple SARS-CoV-2 variants. We demonstrate that mRNA display is an approach for the rapid identification of nAbs that can be used in combination to combat emerging SARS-CoV-2 variants.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Insulin solution human, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
Hemocyanin from Megathura crenulata (keyhole limpet), PBS solution
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KOD Hot Start DNA Polymerase, High fidelity DNA polymerase designed for accurate PCR amplification of long strand and GC- rich DNA templates for cloning and cDNA amplification applications.
Sigma-Aldrich
IgG from human serum, reagent grade, ≥95% (SDS-PAGE), essentially salt-free, lyophilized powder