Protocol for developing Pseudomonas aeruginosa type III secretion system-neutralizing monoclonal antibodies from human B cells

STAR Protoc. 2024 Dec 20;5(4):103440. doi: 10.1016/j.xpro.2024.103440. Epub 2024 Nov 8.

Abstract

Monoclonal antibodies (mAbs) targeting bacterial virulence factors may represent promising therapeutics in the fight against severe bacterial infections. Here, we present an approach for developing human-derived antibodies targeting the type III secretion system (T3SS) of Pseudomonas aeruginosa (PA) by neutralizing the function of the T3SS-tip protein PcrV. The protocol involves identifying individuals with protective antibodies, isolating PcrV-specific B cells from these individuals, and producing and testing anti-PcrV mAbs derived from single B cells. For complete details on the use and execution of this protocol, please refer to Simonis et al.1.

Keywords: biotechnology and bioengineering; cell biology; cell culture; cell isolation; cell-based assays; immunology; microbiology; molecular biology; single cell.

MeSH terms

  • Antibodies, Bacterial / immunology
  • Antibodies, Monoclonal* / immunology
  • Antibodies, Neutralizing* / immunology
  • Antigens, Bacterial / immunology
  • B-Lymphocytes* / immunology
  • B-Lymphocytes* / metabolism
  • Bacterial Proteins / immunology
  • Bacterial Proteins / metabolism
  • Bacterial Toxins
  • Humans
  • Pore Forming Cytotoxic Proteins
  • Pseudomonas Infections / immunology
  • Pseudomonas Infections / microbiology
  • Pseudomonas aeruginosa* / immunology
  • Type III Secretion Systems* / immunology
  • Type III Secretion Systems* / metabolism
  • Virulence Factors / immunology

Substances

  • Type III Secretion Systems
  • Antibodies, Monoclonal
  • Antibodies, Neutralizing
  • Bacterial Proteins
  • Antibodies, Bacterial
  • antigen V, Pseudomonas
  • Antigens, Bacterial
  • Virulence Factors
  • Bacterial Toxins
  • Pore Forming Cytotoxic Proteins