Study of binding and denaturation dynamics of IgG and anti-IgG using dual color fluorescence correlation spectroscopy

Analytica Chimica Acta
Leo Tom VargheseJoseph Irudayaraj

Abstract

In this article, we present a systematic study on IgG and Fab fragment of anti-IgG molecules using fluorescence auto- and cross-correlation spectroscopy to investigate their diffusion characteristics, binding kinetics, and the effect of small organic molecule, urea on their binding. Through our analysis, we found that the diffusion coefficient for IgG and Fab fragment of anti-IgG molecules were 37+/-2 microm2s(-1) and 56+/-2 microm2s(-1), respectively. From the binding kinetics study, the respective forward (k(a)) and backward (k(d)) reaction rates were (5.25+/-0.25)x10(6)M(-1)s(-1) and 0.08+/-0.005 s(-1), respectively and the corresponding dissociation binding constant (K(D)) was 15+/-2 nM. We also found that urea inhibits the binding of these molecules at 4M concentration due to denaturation.

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Citations

Apr 29, 2011·Biosensors & Bioelectronics·Sudeshna PalPhilippe M Fauchet
Mar 31, 2011·Biotechnology and Bioengineering·Brittany BookJoseph Irudayaraj
Mar 22, 2014·Frontiers in Genetics·Yuanyuan ChenFeng C Zhou
Mar 2, 2013·Angewandte Chemie·Nur P DamayantiJoseph M K Irudayaraj
Nov 13, 2013·Optics Express·Jing Liu, Joseph Irudayaraj
Mar 7, 2012·Journal of Cell Science·Jiji ChenJoseph M K Irudayaraj
Sep 7, 2012·The Journal of Physical Chemistry. B·Chaoqing Dong, Joseph Irudayaraj

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