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TitleDevelopment of neutralizing antibodies against SARS-CoV-2, using a high-throughput single-B-cell cloning method.
Journal, issue, pagesAntib Ther, Vol. 6, Issue 2, Page 76-86, Year 2023
Publish dateFeb 17, 2023
AuthorsYang Dou / Ke Xu / Yong-Qiang Deng / Zijing Jia / Jun Lan / Xiaoyu Xu / Guorui Zhang / Tianshu Cao / Pan Liu / Xiangxi Wang / Xinquan Wang / Lingjie Xu / Pan Du / Cheng-Feng Qin / Hong Liu / Yafeng Li / Guizhen Wu / Kang Wang / Bai Lu /
PubMed AbstractBACKGROUND: Rapid and efficient strategies are needed to discover neutralizing antibodies (nAbs) from B cells derived from virus-infected patients.METHODS: Here, we report a high-throughput single-B- ...BACKGROUND: Rapid and efficient strategies are needed to discover neutralizing antibodies (nAbs) from B cells derived from virus-infected patients.
METHODS: Here, we report a high-throughput single-B-cell cloning method for high-throughput isolation of nAbs targeting diverse epitopes on the SARS-CoV-2-RBD (receptor binding domain) from convalescent COVID-19 patients. This method is simple, fast and highly efficient in generating SARS-CoV-2-neutralizing antibodies from COVID-19 patients' B cells.
RESULTS: Using this method, we have developed multiple nAbs against distinct SARS-CoV-2-RBD epitopes. CryoEM and crystallography revealed precisely how they bind RBD. In live virus assay, these nAbs are effective in blocking viral entry to the host cells.
CONCLUSION: This simple and efficient method may be useful in developing human therapeutic antibodies for other diseases and next pandemic.
External linksAntib Ther / PubMed:37077472 / PubMed Central
MethodsEM (single particle)
Resolution3.3 - 6.1 Å
Structure data

31434

7f3q

EMDB-31434, PDB-7f3q:
SARS-CoV-2 RBD in complex with A5-10 Fab and A34-2 Fab
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-31435: SARS-CoV-2 S trimer in complex with 2 A5-10 Fabs (2 RBDs close and 1 RBD open)
Method: EM (single particle) / Resolution: 4.8 Å

EMDB-31436: SARS-CoV-2 S trimer in complex with 1 A5-10 Fab (3 RBDs close)
Method: EM (single particle) / Resolution: 4.7 Å

EMDB-31437: SARS-CoV-2 S trimer in complex with 3 A34-2 Fabs (3 RBDs open)
Method: EM (single particle) / Resolution: 5.3 Å

EMDB-31438: SARS-CoV-2 S trimer in complex with 2 A34-2 Fabs (2 RBDs open and 1 RBD close)
Method: EM (single particle) / Resolution: 5.3 Å

EMDB-31439: SARS-CoV-2 S trimer in complex with 3 A5-10 Fabs (2 RBDs close and 1 RBD open)
Method: EM (single particle) / Resolution: 6.1 Å

Chemicals

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine

ChemComp-MES:
2-(N-MORPHOLINO)-ETHANESULFONIC ACID / pH buffer*YM / MES (buffer)

Source
  • severe acute respiratory syndrome coronavirus 2
  • homo sapiens (human)
KeywordsVIRAL PROTEIN/IMMUNE SYSTEM / SARS-CoV-2 / RBD / Fab / Cocktail / VIRUS / VIRAL PROTEIN-IMMUNE SYSTEM complex

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