Quantitative and qualitative evaluation of adsorption/desorption of bovine serum albumin on hydrophilic and hydrophobic surfaces.
We studied the adsorption of bovine serum albumin (BSA) from phosphate-buffered saline (pH 7.4) to hydrophilic and hydrophobic surfaces. Attenuated total reflection Fourier transform infrared spectroscopy, supported by spectral simulation, allowed us to determine with high precision the amount of BSA adsorbed (surface coverage) and its structural composition. The adsorbed BSA molecules had an alpha-helical structure on both hydrophobic and hydrophilic surfaces but had different molecular conformations and adsorption strengths on the two types of surface. Adsorption of BSA was saturated at around 50% surface coverage on the hydrophobic surface, whereas on the hydrophilic surface the adsorption reached 95%. The BSA molecules adsorbed to the hydrophilic surface with a higher interaction strength than to the hydrophobic surface. Very little adsorbed BSA could be desorbed from the hydrophilic surface, even using 0.1 M sodium dodecyl sulfate, a strong detergent solution. The formation of BSA-phosphate surface complexes was observed under different BSA adsorption conditions on hydrophobic and hydrophilic surfaces. The formation of these complexes correlated with the more efficient blocking of nonspecific interactions by the adsorbed BSA layer. Results from the molecular modeling of BSA interactions with hydrophobic and hydrophilic surfaces support the spectroscopic findings.
Jeyachandran
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YL
LEM, Nancy-Université, CNRS, 15 avenue du Charmois B.P. 40 F-54501 Vandoeuvre lès Nancy, France.
Mielczarski
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E
Rai
B
B
Mielczarski
J A
JA
D000327:Adsorption; D000818:Animals; D002417:Cattle; D005857:Germanium / Q000737:chemistry; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D011137:Polystyrenes / Q000737:chemistry; D011487:Protein Conformation; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared
D011137:Polystyrenes; D005857:Germanium; D012710:Serum Albumin, Bovine