A quantitative and selective chromatography method for determining coverages of multiple proteins on surfaces.
Competitive protein adsorption plays a key role in the surface hemocompatibility of biological implants. We describe a quantitative chromatography method to measure the coverage of multiple proteins physisorbed to surfaces. In this method adsorbed proteins are displaced by CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate) and then analyzed by high performance liquid chromatography to separate and quantify the individual proteins, in this case bovine serum albumin (BSA) and bovine fibrinogen (Fg). CHAPS displaced over 95% of the adsorbed proteins and was easily removed from solution by dialysis. This method was tested by measuring the coverage of BSA, 66 kDa, and Fg, 340 kDa, simultaneously adsorbed from solutions with concentration of 20 microg/ml, on bare and dextranized silicon. Relative to silicon, the dextranized surfaces were found to strongly inhibit protein adsorption, decreasing BSA and Fg coverages by 76 and 60%, respectively.
Ombelli
Michela
M
Department of Anesthesia, University of Pennsylvania, 319B John Morgan Building, 3620 Hamilton Walk, Philadelphia, PA 19104, USA. ombellim@uphs.upenn.edu
Composto
Russell J
RJ
Meng
Qing Cheng
QC
Eckmann
David M
DM
D000327:Adsorption; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D002417:Cattle; D002793:Cholic Acids / Q000737:chemistry; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003911:Dextrans / Q000737:chemistry; D005340:Fibrinogen / Q000737:chemistry; D008422:Materials Testing; D011506:Proteins / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012825:Silicon; D013050:Spectrometry, Fluorescence; D013499:Surface Properties
D001672:Biocompatible Materials; D002793:Cholic Acids; D003911:Dextrans; D011506:Proteins; D012710:Serum Albumin, Bovine; D005340:Fibrinogen; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate; D012825:Silicon