Direct IgG epitope mapping on bacterial AB toxins by cryo-EM

STAR Protoc. 2021 Oct 1;2(4):100852. doi: 10.1016/j.xpro.2021.100852. eCollection 2021 Dec 17.

Abstract

This cryo-EM protocol was used to determine the B cell epitope map on the CdtB subunit of typhoid toxin, an A2B5 toxin secreted by Salmonella Typhi during infection. Immunoglobulin G (IgG) was directly mixed with typhoid toxin in this protocol, different from our previous cryo-EM protocol that uses the Fab fragments in place of IgG. This simple approach requires smaller amounts of materials, supporting the broader use of this protocol for determining antibody recognition sites on various antigens. For complete details on the use and execution of this protocol, please refer to Ahn et al. (2021) and Nguyen et al. (2021).

Keywords: Antibody; Cryo-EM; Immunology; Microbiology; Microscopy; Protein Biochemistry; Structural Biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Toxins*
  • Cryoelectron Microscopy
  • Epitope Mapping
  • Humans
  • Immunoglobulin G
  • Salmonella typhi
  • Typhoid Fever* / microbiology

Substances

  • Bacterial Toxins
  • Immunoglobulin G