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. 2021 Oct;12(10):818-823.
doi: 10.1007/s13238-021-00840-z. Epub 2021 Apr 28.

Molecular deconvolution of the neutralizing antibodies induced by an inactivated SARS-CoV-2 virus vaccine

Xingdong Zhou #  1   2 , Hui Wang #  3 , Qun Ji  2 , Mingjuan Du  2 , Yuexia Liang  1 , Huanhuan Li  1 , Fan Li  1 , Hang Shang  1 , Xiujuan Zhu  3 , Wei Wang  2 , Lichun Jiang  2 , Alexey V Stepanov  4 , Tianyu Ma  1 , Nanxin Gong  1 , Xiaodong Jia  5 , Alexander G Gabibov  4 , Zhiyong Lou  6 , Yinying Lu  7 , Yu Guo  8 , Hongkai Zhang  9   10 , Xiaoming Yang  11
Affiliations

Molecular deconvolution of the neutralizing antibodies induced by an inactivated SARS-CoV-2 virus vaccine

Xingdong Zhou et al. Protein Cell. 2021 Oct.
No abstract available

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Figures

Figure 1
Figure 1
SARS-CoV-2-specific response in human vaccination. (A) Immunization and blood collection regimen. (B) Binding of plasma from donor 1 and donor 2 to SARS-CoV-2 S protein, as determined by ELISA. The mean values and SDs of three technical replicates are shown. (C) Neutralization of two SARS-CoV-2 strains (QD01 and P701) by plasma from donor 1 and donor 2. The mean values and SDs of two technical replicates are shown. (D) Violin plot showing SHM levels (nucleotides) of each donor. The lower, middle and upper edges of the boxplots represent the 25th, 50th and 75th percentiles, respectively. (E) Distribution of heavy chain CDR3 lengths in B cells from vaccinated and naïve donors. (F) Bar graph showing VH germline usage (%) in vaccinated and naïve donors. (G) Outline of microfluidics-based construction of a natively paired VH:VL antibody repertoire. Isolated B cells were purified from blood samples and encapsulated into water-in-oil droplets with beads for mRNA capture. mRNA-captured beads and RT-PCR reagents were reencapsulated, resulting in an amplicon-derived scFv library that can be screened by phage display technology. (H) Schematic of OE-PCR to construct natively paired VH:VL antibody libraries. VH and VL from each encapsulated B cell mRNA are amplified with specific primer sets and paired in-frame via complementary overhangs (yellow). A nested PCR with VH and CL primers generates full-length scFv with SfiI restriction sites for subcloning into phagemid vectors for library generation
Figure 2
Figure 2
Characterization of isolated S protein-reactive antibodies. Sequence information of the identified neutralizing antibodies. 1-0106 is named for clone 1-0106 isolated from donor 1. Midpoint neutralization concentrations (IC50) for four SARS-CoV-2 strains (QD01/P701/HN97/F13). Each symbol represents an individual antibody. A competition assay was performed among different antibodies for binding to the RBD. The immobilized RBD was first incubated with 1-106, 1-108, 2-0126, 2-0139 or 2-01H5. The capture of the second antibodies was monitored by measuring further changes after injecting the second antibody in the presence of the first antibody. (D) Binding of each antibody to RBD and S protein mutants
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References

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