PMID: 3754461Mar 11, 1986Paper

Analysis of protein-protein interaction by simulation of small-zone size-exclusion chromatography: application to an antibody-antigen association

Biochemistry
F J Stevens

Abstract

The association of two or more macromolecules results in the formation of a complex characterized by a larger Stokes radius than that of its components. Therefore, analytical procedures such as ultracentrifugation and size-exclusion gel chromatography that resolve molecules on the basis of size have been used to characterize the association. In this paper we describe an iterative computer simulation of small-zone size-exclusion gel filtration. The simulation describes univalent and bivalent interactions of proteins of equal and nonequal molecular weight and appears to have both qualitative and quantitative application to the evaluation of protein-protein interaction as revealed by alteration of chromatographic elution profiles. To test the validity of the simulation, the model was applied to an antibody-antigen interaction by determining the association constant (Ka) for the interaction between the binding fragment derived from a human immunoglobulin A rheumatoid factor and the antigenic fragment obtained from a human myeloma immunoglobulin G. The self-consistency of the estimated Ka values obtained with a valence value of 2 in contrast to the lack of self-consistency if an antigenic valence of 1 was assumed was taken to suppor...Continue Reading

Citations

Jul 1, 1995·Biomedical Chromatography : BMC·T Cserháti, E Forgács
Apr 6, 1988·Journal of Immunological Methods·F J StevensN S Greenspan
Apr 1, 1990·Journal of Biochemical and Biophysical Methods·F J Stevens
Nov 13, 2009·Journal of the American Chemical Society·Stephan A SchmidLaura M Herz
Jun 15, 2000·Proceedings of the National Academy of Sciences of the United States of America·C T RollinsT Clackson
Feb 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·S MuraoE Huberman
Apr 12, 1994·Proceedings of the National Academy of Sciences of the United States of America·E A MyattF J Stevens
Oct 25, 2000·Protein Science : a Publication of the Protein Society·P R PokkuluriM Schiffer
Dec 25, 2012·Journal of the Mechanical Behavior of Biomedical Materials·C B AbiG Said
May 9, 2006·Biochemical and Biophysical Research Communications·Yinghua ChenOlufunmilayo I Olopade
Dec 30, 2014·Journal of Chromatography. a·Xiao-Shui LiYu-Qi Feng
Jun 15, 1988·Analytical Chemistry·H G BarthF E Regnier
Sep 1, 1995·Protein Science : a Publication of the Protein Society·T P Hopp
Mar 1, 1995·Microbiological Reviews·E M Phizicky, S Fields

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