Immunoaffinity capillary electrophoresis: determination of binding constant and stoichiometry for antibody-antigen interaction

Electrophoresis
Wei-Lung TsengShiming Lin

Abstract

Affinity capillary electrophoresis (ACE) can provide both qualitative and quantitative information on molecular interactions and affords the advantages of very low sample consumption, high mass sensitivity, short analysis time, and the use of automated instrumentation. It has been applied clinically and biochemically to the determination of the binding constant and to the measurement of the binding stoichiometry for interactions between antibodies (Ab's) and antigens (Ag's) in free solution. In many situations, the Ag molecule has two or multiple binding sites, each of which has a similar or different intrinsic affinity for binding independently to the combining site(s) on an Ab molecule. The multivalent binding reactions between Ab and Ag molecules often occur. The objective of this review is to describe the uses of ACE in the determination of binding constants and stoichiometry of Ab-Ag interactions (immunoaffnity capillary electrophoresis), focusing especially on multivalent Ab-Ag interaction modes. Five model binding systems developed recently using ACE techniques are described with principles and examples: (i) divalent mAb-monovalent Ag interaction, (ii) divalent mAb-(homo)polyvalent Ag interaction, (iii) cooperativity of ...Continue Reading

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Citations

May 4, 2005·Analytical Biochemistry·Shiming Lin, Su-Ming Hsu
Nov 25, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·C BertucciV Andrisano
Apr 15, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jian-Feng ZhangYing-Tang Lu
Nov 25, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Carolina SimóAlberto Gallardo
May 31, 2012·ACS Nano·Kiang Wei KhoMalini Olivo
Feb 22, 2014·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Yaghoub SafdariHossein Zarei Jaliani
Jul 25, 2008·Journal of Chromatography. a·Ni LiWenwan Zhong
Dec 15, 2015·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Norberto A Guzman, Daniel E Guzman
Jun 12, 2009·Electrophoresis·Niels H H Heegaard
Dec 7, 2007·Electrophoresis·Carl I D Newman, Greg E Collins
Dec 30, 2009·Electrophoresis·Chunxia Jiang, Daniel W Armstrong
Aug 8, 2015·Journal of Separation Science·Hassan A AlhazmiHermann Wätzig
Feb 1, 2008·Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics·Yousef DaneshbodAli Nadim

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