Citation

  • Authors: Brewer RC. et al.
  • Year: 2022
  • Journal: Nat Immunol 23 33-39
  • Applications: in vitro / DNA / FectoPRO
  • Cell type: Expi293F
    Description: Human embryonic kidney Fibroblast
    Known as: Expi 293-F, Expi, HEK-293 Expi

Method

Recombinant monoclonal antibody production - Heavy chain and light-chain variable sequences were codon optimized and cloned into in-house vectors, containing human IgG1 constant region or κ or λ constant regions, respectively. - Expi293F cells were transfected with heavy chain and light-chain plasmids using FectoPro (Polyplus transfection). - Medium was collected after 7 days and monoclonal antibodies were purified with AmMag Protein A magnetic beads (Genscript).

Abstract

The first ever US Food and Drug Administration-approved messenger RNA vaccines are highly protective against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)1-3. However, the contribution of each dose to the generation of antibodies against SARS-CoV-2 spike (S) protein and the degree of protection against novel variants warrant further study. Here, we investigated the B cell response to the BNT162b2 vaccine by integrating B cell repertoire analysis with single-cell transcriptomics pre- and post-vaccination. The first vaccine dose elicits a recall response of IgA+ plasmablasts targeting the S subunit S2. Three weeks after the first dose, we observed an influx of minimally mutated IgG+ memory B cells that targeted the receptor binding domain on the S subunit S1 and likely developed from the naive B cell pool. This response was strongly boosted by the second dose and delivers potently neutralizing antibodies against SARS-CoV-2 and several of its variants.

Pubmed