Comparison of covalent and noncovalent labeling with near-infrared dyes for the high-performance liquid chromatographic determination of human serum albumin.
Noncovalent and covalent methods of labeling protein with near-infrared polymethine cyanine dyes were compared for use in analyzing human serum albumin (HSA) by high-performance liquid chromatography (HPLC) with near-infrared absorbance detection. While noncovalent labeling was faster than covalent labeling and took place in the physiological pH range, covalent labeling was more stable under conditions encountered in many of the widely used types of HPLC. Covalently labeled HSA protein peaks indicated uniform labeling of amino groups at both hydrophilic and hydrophobic binding sites, while noncovalent labeling showed a preference for hydrophobic binding sites.
Williams
R J
RJ
Department of Chemistry, Georgia State University, Atlanta 30303.
Lipowska
M
M
Patonay
G
G
Strekowski
L
L
D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D012709:Serum Albumin / Q000032:analysis*; D013055:Spectrophotometry, Infrared
D005456:Fluorescent Dyes; D012709:Serum Albumin