Comparative analysis of two meningococcal immunotyping monoclonal antibodies by resonant mirror biosensor and antibody gene sequencing.
Lipooligosaccharide (LOS) is a major surface component of the cell walls of Neisseria meningitidis, which is important for its roles in pathogenesis and antigenic variation, as a target for immunological typing, and as a possible vaccine component. Although the structures of many antigenic variants have been determined, routine immunological typing of these molecules remains problematic. Resonant mirror analysis was combined with gene sequencing to characterize two monoclonal antibodies (MAbs) used in typing panels that were raised against the same LOS immunotype, L3,7,9. The two MAbs (MAb 4A8-B2 and MAb 9-2-L379) were of the same immunoglobulin subtype, but while MAb 9-2-L379 was more than a 1,000-fold more sensitive in immunotyping assays of both whole meningococcal cells and purified LOS, MAb 4A8-B2 was more specific for immunotype L3,7,9. The differences in sensitivity were a consequence of MAb 9-2-L379 having a 44-fold-faster association constant than MAb 4A8-B2. Comparison of the amino acid sequences of the variable chains of the MAbs revealed that they had very similar heavy chains (81% amino acid sequence identity) but diverse light chains (54% sequence identity). The differential binding kinetics and specificities observed with these MAbs were probably due to differences in the epitopes recognized, and these were probably a consequence of the different immunization protocols used in their production.
Charalambous
B M
BM
Department of Biochemistry and Molecular Biology, Royal Free, United Kingdom.
Evans
J
J
Feavers
I M
IM
Maiden
M C
MC
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D001667:Binding, Competitive / Q000276:immunology; D015374:Biosensing Techniques; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D007114:Immunization; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D007700:Kinetics; D008585:Meningitis, Meningococcal / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000276:immunology*; D017422:Sequence Analysis, DNA; D013745:Tetanus Toxoid / Q000276:immunology / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D007135:Immunoglobulin Variable Region; D013745:Tetanus Toxoid