Nolles Antsje, Westphal Adrie H, de Hoop Jacob A, Fokkink Remco G, Kleijn J Mieke, van Berkel Willem J H, Borst Jan Willem
†Laboratory of Biochemistry, ‡Physical Chemistry and Soft Matter, and §MicroSpectroscopy Centre Wageningen, Wageningen University, Wageningen, The Netherlands.
Biomacromolecules. 2015 May 11;16(5):1542-9. doi: 10.1021/acs.biomac.5b00092. Epub 2015 Apr 16.
Protein encapsulation with polymers has a high potential for drug delivery, enzyme protection and stabilization. Formation of such structures can be achieved by the use of polyelectrolytes to generate so-called complex coacervate core micelles (C3Ms). Here, encapsulation of enhanced green fluorescent protein (EGFP) was investigated using a cationic-neutral diblock copolymer of two different sizes: poly(2-methyl-vinyl-pyridinium)41-b-poly(ethylene-oxide)205 and poly(2-methyl-vinyl-pyridinium)128-b-poly(ethylene-oxide)477. Dynamic light scattering and fluorescence correlation spectroscopy (FCS) revealed a preferred micellar composition (PMC) with a positive charge composition of 0.65 for both diblock copolymers and micellar hydrodynamic radii of approximately 34 nm. FCS data show that at the PMC, C3Ms are formed above 100 nM EGFP, independent of polymer length. Mixtures of EGFP and nonfluorescent GFP were used to quantify the amount of GFP molecules per C3M, resulting in approximately 450 GFPs encapsulated per micelle. This study shows that FCS can be successfully applied for the characterization of protein-containing C3Ms.
用聚合物包裹蛋白质在药物递送、酶保护和稳定方面具有很高的潜力。通过使用聚电解质来生成所谓的复合凝聚核胶束（C3M），可以实现这种结构的形成。在此，使用两种不同尺寸的阳离子-中性二嵌段共聚物：聚（2-甲基-乙烯基吡啶鎓）41-b-聚（环氧乙烷）205和聚（2-甲基-乙烯基吡啶鎓）128-b-聚（环氧乙烷）477，研究了增强型绿色荧光蛋白（EGFP）的包裹情况。动态光散射和荧光相关光谱（FCS）显示，两种二嵌段共聚物都具有一个优选的胶束组成（PMC），其正电荷组成为0.65，胶束流体动力学半径约为34 nm。FCS数据表明，在PMC下，当EGFP浓度高于100 nM时会形成C3M，与聚合物长度无关。使用EGFP和非荧光GFP的混合物来量化每个C3M中GFP分子的数量，结果是每个胶束中约包裹450个GFP。这项研究表明，FCS可以成功地应用于含蛋白质C3M的表征。
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