Partial filling affinity capillary electrophoresis with cationic poly(vinylpyrrolidone)-based copolymer coatings for studies on human lipoprotein-steroid interactions

作者：Wang Ai Jun; Vainikka Kati; Witos Joanna; D'Ulivo Lucia; Cilpa Geraldine; Kovanen Petri T; Oorni Katariina; Jauhiainen Matti; Riekkola Marja Liisa^*

来源：Analytical Biochemistry, 2010, 399(1): 93-101.

DOI：10.1016/j.ab.2009.11.028

• Affinity capillary electrophoresis
• Partial filling technique
• Affinity constant
• High-and low-density lipoproteins
• Steroids
• Copolymers

Human plasma lipoproteins have strong hydrophobic interactions with steroids and their fatty acyl derivatives such as estradiol fatty acyl esters. In this work, affinity capillary electrophoresis with the partial filling technique was applied to study the hydrophobic interactions between lipoproteins, which are nanometer-sized particles, and nonconjugated steroids. The capillaries were first rinsed with one of two novel poly(vinylpyrrolidone) (PVP)-based cationic copolymers that were strongly adsorbed onto the fused-silica surface via electrostatic interactions. This surface treatment greatly suppressed the adsorption of lipoproteins. Low-density lipoprotein (LDL) and high-density lipoprotein (HDL) particles were then employed in the coated capillaries as pseudostationary phase in the partial filling mode. The changes in corrected migration times of steroids increased linearly with the filling time of the lipoproteins. The affinity constants between the steroids and lipoproteins were calculated, and the most hydrophobic steroid studied, progesterone, had stronger affinity than testosterone or androstenedione toward both LDL and HDL. Affinity between steroids and LDL was stronger than that between steroids and HDL. Interactions between the steroids and lipoproteins were mainly nonspecific with particle lipid components, whereas some were site specific with the apolipoproteins. The developed technique has great potential for determination of the affinity of various compounds toward lipoproteins.

• 出版日期2010-4-1