Project 2: Nanobodies as Novel Entry Inhibitors of Pandemic Viruses

**Construction of a camelid nanobody phage display library and use of this library for screening of anti-SARS-CoV-2 nanobodies.**

A large-sized (diversity 7.5 x 1010) naive nanobody phage display library was constructed using B cells of over a dozen llamas and alpacas. Phages were screened for their high binding affinity for SARS-CoV-2 receptor-binding domain (RBD). Nanobodies expressed from the selected phages were further screened for their potency in neutralizing SARS-CoV-2 pseudovirus entry. The best performing nanobody was subjected to two rounds of affinity maturation.

Project Description

The fast emergence of many SARS-CoV-2 variants calls for the urgent need for potent and broad-spectrum anti-COVID-19 therapeutics. Compared to small-molecule antiviral drugs, nanobodies are particularly powerful in battling SARS-CoV-2 variants because they can be quickly adapted to new viral variants.

Moreover, other pathogenic viruses also demonstrate pandemic potential, such as Ebola, Lassa and Zika viruses. These RNA viruses all contain a surface glycoprotein (a molecule comprised of protein and carbohydrate chains) that mediates virus entry into host cells; thus, the viral glycoprotein serves as a critical therapeutic target.

Project 2 proposes to develop highly effective nanobodies as novel inhibitors to block the viral entry of these pandemic viruses. In our prior studies, we have developed several potent nanobodies, including a series of anti-SARS-CoV-2 candidate drugs named Nanosota-1.

We have established a camelid nanobody phage display for screening. Nanobodies with high potency, good stability, low production costs, minimal side effects, and a broad antiviral spectrum can be developed as novel antiviral therapeutics.

Principal Investigator

Lanying Du, Ph.D: Georgia State University, lab leak, covid-19, wuhan — Lanying Du, Ph.D

[email protected]

Georgia State University, UT San Antonio Health, University of Iowa, Iowa State University, Nanyang technical university, University of Illinois Chicago, University of Louisville, University of Tennessee, university of Minnesota, UC Berkeley, UC San Diego, UF Scripps Biomedical Research, Baylor, Boston University, New York Blood Center,

Deputy Investigator

Fang Li, Ph.D: University of Minnesota, coronavirus research center, spike protein, Midwest AViDD, University of minnesota, llama, lab leak, wuhan, — **Fang Li, Ph.D**

*[email protected]*

Deputy Investigator

Michael Farzan, UF Scripps Biomedical Research, Lab Leak, Covid-19, Project 1: Small Molecule Entry Inhibitors of Pandemic Viruses — ***Michael Farzan, Ph.D***

[email protected]

Choe Hyeryun, Project 1: Small Molecule Entry Inhibitors of Pandemic Viruses, UF Scripps Biomedical Research — **Hyeryun Choe, Ph.D: UF Scripps**

*[email protected]*

Stanley Perlman, M.D, Ph.D: University of Iowa, OSC, nanobody, nanosota-1, camelid, llama, sars, covid-19 — **Stanley Perlman, M.D, Ph.D: University of Iowa, OSC**

[email protected]

Matthew Aliota, Ph.D: University of Minnesota, OSC, aliota lab, animal testing, viruses, pandemic, virology, mutations — **Matthew Aliota, Ph.D: University of Minnesota, OSC**

[email protected]

Lijun Rong, Ph.D: University of Illinois Chicago, OSC, pandemic, virus, covid-19, antiviral, midwest avidd — **Lijun Rong, Ph.D: University of Illinois Chicago, OSC**

[email protected]

Potent Nanobodies

Principal Investigator

Lanying Du, Ph.D

Deputy Investigator

Deputy Investigator

Co-Investigators and Other Significant Contributors (OSC)