Opsonophagocytic Killing Assay to Measure Anti-Group A Streptococcus Antibody Functionality in Human Serum

Helen Wagstaffe; Scott Jones; Marina Johnson; David Goldblatt

doi:10.1007/978-1-0716-1900-1_20

Opsonophagocytic Killing Assay to Measure Anti-Group A Streptococcus Antibody Functionality in Human Serum

Methods Mol Biol. 2022:2414:373-386. doi: 10.1007/978-1-0716-1900-1_20.

Authors

Helen Wagstaffe^{1

2}, Scott Jones³, Marina Johnson³, David Goldblatt³

Affiliations

¹ Great Ormond Street Institute of Child Health, University College London, London, UK. [email protected].
² Department of Infectious Disease, Imperial College London, London, UK. [email protected].
³ Great Ormond Street Institute of Child Health, University College London, London, UK.

PMID: 34784047
DOI: 10.1007/978-1-0716-1900-1_20

Abstract

The opsonophagocytic killing assay (OPKA) is designed to measure the functionality of strain-specific antibodies and, therefore, assess protective immunity or the immunogenicity of Group A Streptococcus (GAS) (type A Streptococcus pyogenes) vaccines. Opsonization of GAS for phagocytosis is an important mechanism by which antibodies protect against disease in vivo. The Opsonophagocytic Index or Opsonic Index (OI) is the estimated dilution of antisera that kills 50% of the target bacteria. Here, we describe the protocol of the standardized GAS OPKA developed by Jones et al., 2018.

Keywords: Antibody; Baby rabbit complement; Functional assay; HL-60 cells; In vitro; Opsonophagocytic killing assay; Phagocytosis; S. pyogenes.

MeSH terms

Antibodies, Bacterial
Biological Assay
Humans
Opsonin Proteins
Phagocytosis
Streptococcus pyogenes* / immunology

Substances

Antibodies, Bacterial
Opsonin Proteins