On the interaction between a bactericidal antibody and a PorA epitope of Neisseria meningitidis in outer membrane vesicles: a competitive fluorescence polarization immunoassay

Analytical Biochemistry
J M van den ElsenD J Crommelin

Abstract

This paper describes a method for determining the affinity constant (Ka) of the binding between an antibody Fab fragment and a membrane-embedded protein epitope under equilibrium conditions. Monoclonal antibody MN12H2, directed against outer membrane protein PorA of Neisseria meningitidis, is used in a competitive fluorescence polarization assay with a cyclic peptide-fluorescein conjugate as a tracer antigen. Displacement experiments with PorA-containing and PorA-deficient meningococcal outer membrane vesicles revealed highly specific binding of MN12H2 Fab to the membrane-embedded PorA P1.16 epitope with Ka of 1.5 x 10(8) M-1.

References

Sep 1, 1991·Infection and Immunity·P van der LeyJ T Poolman
Apr 25, 1991·Journal of Immunological Methods·W JiskootE C Beuvery
Dec 1, 1990·Pharmaceutical Research·W JiskootD J Crommelin
Aug 1, 1989·Immunology Today·M H Van Regenmortel
Nov 1, 1973·The Journal of Experimental Medicine·I W Devoe, J E Gilchrist
Jan 1, 1982·Methods in Enzymology·B J Litman, Y Barenholz

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Citations

Oct 1, 2005·Protein Science : a Publication of the Protein Society·Zhiwen XuHong Xue
Dec 1, 2006·Molecular Biology and Evolution·Shui-Ying TsangHong Xue

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