Adsorption of bovine serum albumin (BSA) onto lecithin studied by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy.
The adsorption of bovine serum albumin (BSA) to lecithin was investigated by ATR-FTIR spectroscopy. Lecithin films were prepared by casting aliquots of 3.2 microg lecithin in methanol onto ZnSe ATR prisms. Surface morphology and the thickness of the films were investigated by laser scanning confocal electron microscopy and scanning electron microscopy and the thickness of the films used for adsorption studies was estimated to be 40 A. The dependency of the CO peak area on the lecithin mass in the calibration curve confirms that the thickness of the film is below the penetration depth of the infrared evanescent wave. Size exclusion HPLC and fluorescence spectroscopy show that BSA conformation in up to 1M NaCl and CaCl(2) solutions is similar to that in water with no aggregation or changes in protein conformation seen over 4h. The kinetics of BSA adsorption on the lecithin film from water, NaCl and CaCl(2) solutions demonstrates that ions promote the protein adsorption. BSA bound more in the presence of NaCl compared to CaCl(2) at equivalent concentrations. The adsorption appeared greatest at a 0.1M concentration for both NaCl and CaCl(2). The results are explained in terms of absorptive reactivity of BSA and lecithin surfaces upon salt addition.
Tantipolphan
R
R
School of Pharmacy, The University of Otago, Dunedin, New Zealand.
Rades
T
T
McQuillan
A J
AJ
Medlicott
N J
NJ
D000327:Adsorption; D000818:Animals; D002122:Calcium Chloride / Q000737:chemistry*; D002417:Cattle; D002626:Chemistry, Pharmaceutical; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid; D005079:Excipients / Q000737:chemistry*; D007700:Kinetics; D000432:Methanol / Q000737:chemistry; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D010713:Phosphatidylcholines / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D018036:Selenium Compounds / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism; D012965:Sodium Chloride / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared; D013499:Surface Properties; D013678:Technology, Pharmaceutical / Q000379:methods*; D014867:Water / Q000737:chemistry; D017967:Zinc Compounds / Q000737:chemistry
D005079:Excipients; D010713:Phosphatidylcholines; D018036:Selenium Compounds; D017967:Zinc Compounds; D014867:Water; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D002122:Calcium Chloride; C044696:zinc selenide; D000432:Methanol