Recombinant Albumin Monolayers on Latex Particles

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• 作者：Kamila Sofińska ; Zbigniew Adamczyk ; Marta Kujda ; Ma?gorzata Nattich-Rak
• 刊名：Langmuir
• 出版年：2014
• 出版时间：January 14, 2014
• 年：2014
• 卷：30
• 期：1
• 页码：250-258
• 全文大小：419K
• ISSN：1520-5827

文摘

The adsorption of recombinant human serum albumin (rHSA) on negatively charged polystyrene latex micro-particles was studied at pH 3.5 and the NaCl concentration range of 10^鈥? to 0.15 M. The electrophoretic mobility of latex monotonically increased with the albumin concentration in the suspension. The coverage of adsorbed albumin was quantitatively determined using the depletion method, where the residual protein concentration was determined by electrokinetic measurements and AFM imaging. It was shown that albumin adsorption was irreversible. Its maximum coverage on latex varied between 0.7 mg m^鈥? for 10^鈥? M NaCl to 1.3 mg m^鈥? for 0.15 M NaCl. The latter value matches the maximum coverage previously determined for human serum albumin on mica using the streaming potential method. The increase in the maximum coverage was interpreted in terms of reduced electrostatic repulsion among adsorbed molecules. These facts confirm that albumin adsorption at pH 3.5 is governed by electrostatic interactions and proceeds analogously to colloid particle deposition. The stability of albumin monolayers was measured in additional experiments where changes in the latex electrophoretic mobility and the concentration of free albumin in solutions were monitored over prolonged time periods. Based on these experimental data, a robust procedure of preparing albumin monolayers on latex particles of well-controlled coverage and molecule distribution was proposed.