Adsorption of bovine serum albumin on fused silica: Elucidation of protein-protein interactions by single-molecule fluorescence microscopy

Colloids and Surfaces. B, Biointerfaces
K M YeungN H Cheung

Abstract

The adsorption of bovine serum albumin (BSA) on fused silica at pH 4.7 was studied at the single molecules level by total-internal-reflection fluorescence microscopy. This pH value was the isoelectric point of BSA. At low [BSA] of 20pM, protein molecules adsorbed as monomers. At intermediate [BSA] of 500pM, protein molecules adsorbed as clusters of about five monomers on average. Both monomers and clusters had adsorption rate coefficients of the order 10(-7)ms(-1) and desorption rate coefficients of about 2x10(-2)s(-1). The respective steady-state coverage was about 10x higher than that at neutral pH, presumably because of the more favorable BSA-silica electrostatics. At pH 4.7 and with [BSA] higher than 100nM, adsorption begot further adsorption to produce nonlinear isotherms. The coverage at 1microM BSA was 2.5x that of the linearly extrapolated coverage. This suggests that at pH 4.7, solute-adsorbate affinity was the dominant factor that explains the enhanced adsorption observed in ensemble measurements.

References

Dec 19, 2006·Journal of Colloid and Interface Science·Laura E ValentiCarla E Giacomelli
Nov 21, 2007·Colloids and Surfaces. B, Biointerfaces·Willem Norde

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Citations

Jun 22, 2012·Langmuir : the ACS Journal of Surfaces and Colloids·Mark KastantinDaniel K Schwartz
Feb 4, 2014·Advances in Colloid and Interface Science·Mark KastantinDaniel K Schwartz
Dec 9, 2009·Colloids and Surfaces. B, Biointerfaces·Elena PrimoracDavor Kovacević
May 8, 2009·Colloids and Surfaces. B, Biointerfaces·Guillaume LamourAhmed Hamraoui
Mar 12, 2016·International Journal of Biological Macromolecules·Sayyid Mahdi RafighHamid Mozafari
Feb 8, 2011·Advances in Colloid and Interface Science·Michael RabeStefan Seeger
Aug 29, 2012·Analytical Chemistry·Thomas C Oates, Lloyd W Burgess
Oct 30, 2016·International Journal of Biological Macromolecules·Abid HussainMohammad Zuber

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