Properties of Monoclonal Antibodies to the Genus-specific Antigen of Chlamydia and Their Use for Antigen Detection by Reverse Passive Haemagglutination

• Published in: Journal of general microbiology Vol. 131; no. 1; pp. 7 - 15
• Main Authors: THORNLEY, MARGARET J, ZAMZE, SUSANNE E, BYRNE, MARIE D, LUSHER, MERYL, EVANS, ROGER T
• Format: Journal Article
• Language: English
• Published: London Soc General Microbiol 01.01.1985; New York, NY Cambridge University Press
• Subjects: Antibodies, Monoclonal; Antigens, Bacterial; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Biotechnology; Chlamydia - immunology; Chlamydia trachomatis - immunology; Chlamydophila psittaci - immunology; Cross Reactions; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Fundamental and applied biological sciences. Psychology; Health. Pharmaceutical industry; Hemagglutination Tests; Immunodiffusion; Immunoglobulin G; Industrial applications and implications. Economical aspects; Microbiology; Monoclonal antibodies; Production of active biomolecules; Antigen; Hemagglutination; Chlamydiaceae; Chlamydia; Chlamydiales; Lipopolysaccharide; Bacteria; Monoclonal antibody; Detection; ELISA assay; Immunological method
• DOI: 10.1099/00221287-131-1-7
• ISSN: 0022-1287; 1350-0872

Division of Immunology, Department of Pathology, University of Cambridge, Addenbrooke's Hospital, Cambridge CB2 2QQ, UK Division of Sexually Transmitted Diseases, Clinical Research Centre, Watford Road, Harrow, Middlesex HA1 3UJ, UK ABSTRACT SUMMARY: The chlamydial genus-specific antigen was extracted with phenol/chloroform petroleum ether (PCP) from preparations of Chlamydia trachomatis and C. psittaci , and quantities measured using an assay for lipopolysaccharide (LPS). The LPS from C. trachomatis contained 2·2% (w/w) of ketodeoxyoctanoic acid. Five IgG monoclonal antibodies reacted in an ELISA with LPS from both species, the antigen being periodate-sensitive and heat-resistant, confirming that all antibodies were against the genus-specific antigen. All the antibodies bound to the PCP extract of C. trachomatis on an immunoblot, at a position corresponding to the periodate-Schiff-stained bands of both C. trachomatis extract and Salmonella Re-LPS. When linked to trypsin-treated sheep erthrocytes and used in reverse passive haemagglutination tests, all antibodies gave indicator cells capable of detecting chlamydial LPS or crude preparations of chlamydiae grown in McCoy cells, the sensitivity varying with the antibody used. The antibodies varied in IgG subclass (either IgG 2a or IgG 3 ), and in ability to precipitate in immunodiffusion tests. Two antibodies cross-reacted with one strain of Acinetobacter in ELISA and with Salmonella Re-LPS in both ELISA and immunodiffusion tests. The other three did not react in ELISA with Acinetobacter strains or Salmonella Re-LPS, and none of the five reacted with LPS of E. coli or Pseudomonas morsprunorum . Present address: Department of Paediatrics, John Radcliffe Hospital, Headington, Oxford, OX3 9DU, UK.