应用FRAP技术评价蛋白在聚丙烯酰胺-丙烯酸凝胶中的扩散行为

Study on the Protein Diffusion Behavior in the Poly(Acrylamide-co-Acrylic Acid) Copolymer by FRAP Technique

药物分子从药物载体中的释放行为与载体的结构有密切关系.本实验中采用丙烯酰胺和丙烯酸等材料,运用水相沉淀的方法,制备了4种不同单体配比的聚丙烯酰胺-丙烯酸(P(Am-co-Ac))共聚物水凝胶.运用红外分析方法对P(Am-co-Ac)组成进行表征.使用荧光漂白恢复法(FRAP,fluorescence recovery after photobleaching)观察荧光素FITC标记的牛血清白蛋白(BSA)在聚丙烯酰胺-丙烯酸中的扩散行为,并以激光共聚焦显微镜进行实时成像.实验表明,FITC-BSA在不同单体配比的共聚物中的扩散系数是不同的.通过调节聚合物中单体的配比能够达到控制蛋白释放速率的作用,从而为调控蛋白和多肽类药物的缓控释放提供了可能性.

The drug release behavior from the carrier is bound up with the carrier＇s structure. In this paper, a series of poly （acrylamide-co-acrylic acid） hydrogels with different acrylamide/ acrylic acid ratios have been successfully synthesized by aqueous polymerization. These co-polymers were characterized with FTIR technique to confirm the co-polymer formation. Fluorescence recovery after photobleaching （FRAP） technique was used to investigate the diffusion behavior of fluorescein isothiocyanate labelled bovine serum albumin （FITC-BSA） in these co-polymer hydrogels, and a confocal laser scanning microscopy was applied to obtain high resolution images in real time. The results showed that the diffusion coefficient of FITCBSA was influenced by the ratio between the acrylamide and acrylic acid in co-polymer and the co-polymer hydrogels synthesized may have potential application in controlled protein delivery system.