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Protocol for constructing large size human antibody heavy chain variable domain (V_H) library and selection of SARS-CoV-2 neutralizing antibody domains.

ABSTRACT: This protocol is a comprehensive guide to phage display-based selection of virus neutralizing V_H 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 V_H antibody domain, V_H 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).

SUBMITTER: Chen C

PROVIDER: S-EPMC8164376 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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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 C Sun Zehua Z Liu Xianglei X Li Wei W Dimitrov Dimiter S DS

STAR protocols 20210529 3

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 hu ...[more]

PMID: 34095859

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