ON THE INTERACTION BETWEEN A BACTERICIDAL ANTIBODY AND A PORA EPITOPEOF NEISSERIA-MENINGITIDIS IN OUTER-MEMBRANE VESICLES - A COMPETITIVE FLUORESCENCE POLARIZATION IMMUNOASSAY

Authors
VANDENELSEN JMH VANPOMEREN E POOLMAN JT WILTING J HERRON JN CROMMELIN DJA
Citation
Jmh. Vandenelsen et al., ON THE INTERACTION BETWEEN A BACTERICIDAL ANTIBODY AND A PORA EPITOPEOF NEISSERIA-MENINGITIDIS IN OUTER-MEMBRANE VESICLES - A COMPETITIVE FLUORESCENCE POLARIZATION IMMUNOASSAY, Analytical biochemistry, 247(2), 1997, pp. 382-388
Citations number
27
Categorie Soggetti
Biology
Journal title
Analytical biochemistry → ACNP
ISSN journal
00032697
Volume
247
Issue
2
Year of publication
1997
Pages
382 - 388
Database
ISI
SICI code
0003-2697(1997)247:2<382:OTIBAB>2.0.ZU;2-7
Abstract
This paper describes a method for determining the affinity constant (K -a) of the binding between an antibody Fab fragment and a membrane-emb edded protein epitope under equilibrium conditions. Monoclonal antibod y MN12H2, directed against outer membrane protein PorA of Neisseria me ningitidis, is used in a competitive fluorescence polarization assay w ith a cyclic peptide-fluorescein conjugate as a tracer antigen. Displa cement experiments with PorA-containing and PorA-deficient meningococc al outer membrane vesicles revealed highly specific binding of MN12H2 Fab to the membrane embedded PorA P1.16 epitope with K-a of 1.5 x 10(8 ) M-1. (C) 1997 Academic Press.