Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains
Summary: 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 e...
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2021-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721003245 |
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doaj-397aa765dacb430796964e1a527673992021-09-19T04:59:55ZengElsevierSTAR Protocols2666-16672021-09-0123100617Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domainsChuan Chen0Zehua Sun1Xianglei Liu2Wei Li3Dimiter S. Dimitrov4Center for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USA; Corresponding authorCenter for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USACenter for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USACenter for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USA; Corresponding authorCenter for Antibody Therapeutics, Division of Infectious Diseases, Department of Medicine, University of Pittsburgh Medical School, Pittsburgh, PA, USA; Abound Bio, Pittsburgh, PA, USA; Corresponding authorSummary: 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)http://www.sciencedirect.com/science/article/pii/S2666166721003245Cell cultureHigh Throughput ScreeningImmunologyMicrobiologyMolecular BiologyAntibody |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Chuan Chen Zehua Sun Xianglei Liu Wei Li Dimiter S. Dimitrov |
| spellingShingle |
Chuan Chen Zehua Sun Xianglei Liu Wei Li Dimiter S. Dimitrov Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains STAR Protocols Cell culture High Throughput Screening Immunology Microbiology Molecular Biology Antibody |
| author_facet |
Chuan Chen Zehua Sun Xianglei Liu Wei Li Dimiter S. Dimitrov |
| author_sort |
Chuan Chen |
| title |
Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains |
| title_short |
Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains |
| title_full |
Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains |
| title_fullStr |
Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains |
| title_full_unstemmed |
Protocol for constructing large size human antibody heavy chain variable domain (VH) library and selection of SARS-CoV-2 neutralizing antibody domains |
| title_sort |
protocol for constructing large size human antibody heavy chain variable domain (vh) library and selection of sars-cov-2 neutralizing antibody domains |
| publisher |
Elsevier |
| series |
STAR Protocols |
| issn |
2666-1667 |
| publishDate |
2021-09-01 |
| description |
Summary: 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) |
| topic |
Cell culture High Throughput Screening Immunology Microbiology Molecular Biology Antibody |
| url |
http://www.sciencedirect.com/science/article/pii/S2666166721003245 |
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