Covalent immobilization of human serum albumin onto reactive polypyrrole-coated polystyrene latex particles
Covalent immobilization of human serum albumin onto reactive polypyrrole-coated polystyrene latex particles
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DOI：
10.1039/b500982k
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发表时间：
2005-01-01
影响因子：
--
通讯作者：
Chehimi, MM
中科院分区：
文献类型：
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作者：
Bousalem, S;Benabderrahmane, S;Chehimi, MM
在直径为1 mm的聚苯乙烯（PS）乳胶粒存在下，通过吡咯（Py）和活性酯官能化吡咯（PyNSE）的原位共聚反应制备了带有反应性N-琥珀酰亚胺基官能团的聚吡咯包覆聚苯乙烯乳胶粒（PS-PPyNSE）。这些粒子是通过使用聚（N-乙烯基吡咯烷酮），PNVP，作为空间稳定剂的苯乙烯的分散聚合制备的。对于吡咯和吡咯-NSE，初始共聚单体分数（以摩尔%计）分别为25/75、50/50和75/25。功能化的聚吡咯包覆的PS颗粒（PS-PPyNSE），其特征在于在其粒径和表面形态，使用透射电子显微镜（TEM）。红外和X射线光电子能谱（XPS）检测到吡咯-NSE重复单元在乳胶粒子的表面，表明该单体确实与吡咯共聚。此外，PS-PPyNSE颗粒的核-壳结构通过在THF中蚀刻聚苯乙烯核来确认，导致中空导电聚合物胶囊的形成。PS-PPyNSE 75颗粒的反应性使用2-氨基乙醇和2-巯基乙醇进行了研究，这两种模型分子带有由蛋白质携带的官能团。将颗粒与这些模型分子一起孵育清楚地表明，颗粒对胺和硫醇基团具有反应性，分别导致界面酰胺和硫酯键的形成。进一步评估PS-PPyNSE颗粒作为用作测试蛋白的人血清白蛋白（HSA）的生物吸附剂。结果表明，HSA大分子通过形成界面酰胺基团固定在PS-PPyNSE颗粒的表面。HSA接枝的胶乳颗粒与抗HSA的孵育导致立即絮凝。这表明，尽管与NSE-官能化胶乳接触可能导致构象变化，但HSA蛋白在固定在颗粒表面上时保留其生物活性。因此，这些PS-PPyNSE颗粒是视觉诊断测定的替代候选物。
Polypyrrole-coated polystyrene latex particles bearing reactive N-succinimidyl functional groups (PS-PPyNSE) were prepared by the in-situ copolymerization of pyrrole (Py) and the active ester-functionalized pyrrole (PyNSE) in the presence of 1 mm diameter-sized polystyrene ( PS) latex particles. These particles were prepared by dispersion polymerization of styrene using poly(N-vinylpyrrolidone), PNVP, as a steric stabilizer. The initial comonomer fractions (in mol%) were 25/75, 50/50 and 75/25 for pyrrole and pyrrole-NSE, respectively. The functionalized polypyrrole-coated PS particles (PS-PPyNSE) were characterized in terms of their particle size and surface morphology using transmission electron microscopy (TEM). Infrared and X-ray photoelectron spectroscopy (XPS) detected pyrrole-NSE repeat units at the surface of the latex particles, indicating that this monomer had indeed copolymerized with pyrrole. Furthermore, the core-shell structure of the PS-PPyNSE particles was confirmed by etching the polystyrene core in THF, leading to the formation of hollow conducting polymer capsules. Reactivity of the PS-PPyNSE75 particles was investigated using 2-aminoethanol and 2-mercaptoethanol, two model molecules bearing functional groups borne by proteins. Incubation of the particles with these model molecules clearly showed that the particles are reactive towards amine and thiol groups leading to the formation of interfacial amide and thioester bonds, respectively. The PS-PPyNSE particles were further evaluated as bioadsorbents of human serum albumin (HSA) used as a test protein. It was shown that the HSA macromolecules were immobilized at the surface of PS-PPyNSE particles by forming interfacial amide groups. Incubation of the HSA-grafted latex particles with anti-HSA resulted in immediate flocculation. This indicates that, despite probable conformational changes resulting from contact with NSE-functionalized latexes, HSA proteins retain their biological activity when immobilized on the particle surface. These PS-PPyNSE particles are therefore alternative candidates for visual diagnostic assays.