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Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains.
Chen, Chuan; Sun, Zehua; Liu, Xianglei; Li, Wei; Dimitrov, Dimiter S.
  • Chen C; Center for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USA.
  • Sun Z; Center for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USA.
  • Liu X; Center for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USA.
  • Li W; Center for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USA.
  • Dimitrov DS; Center for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USA.
STAR Protoc ; 2(3): 100617, 2021 09 17.
Article in English | MEDLINE | ID: covidwho-1386745
ABSTRACT
This protocol is a comprehensive guide to phage display-based selection of virus neutralizing VH antibody domains. It details three optimized parts including (1) construction of a large-sized (theoretically > 1011) naïve human antibody heavy chain domain library, (2) SARS-CoV-2 antigen expression and stable cell line construction, and (3) library panning for selection of SARS-CoV-2-specific antibody domains. Using this protocol, we identified a high-affinity neutralizing human VH antibody domain, VH ab8, which exhibits high prophylactic and therapeutic efficacy. For complete details on the use and execution of this protocol, please refer to Li et al. (2020).
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Immunoglobulin Variable Region / Immunoglobulin Heavy Chains / Peptide Library / Antibodies, Neutralizing / SARS-CoV-2 / COVID-19 / Antibodies, Viral Type of study: Randomized controlled trials Limits: Humans Language: English Journal: STAR Protoc Year: 2021 Document Type: Article Affiliation country: J.XPRO.2021.100617

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Immunoglobulin Variable Region / Immunoglobulin Heavy Chains / Peptide Library / Antibodies, Neutralizing / SARS-CoV-2 / COVID-19 / Antibodies, Viral Type of study: Randomized controlled trials Limits: Humans Language: English Journal: STAR Protoc Year: 2021 Document Type: Article Affiliation country: J.XPRO.2021.100617