Preparation of multilayer plasma protein films on silicon by EDC/NHS coupling chemistry
Preparation of multilayer plasma protein films on silicon by EDC/NHS coupling chemistry
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DOI：
10.1016/s0927-7765(02)00151-0
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发表时间：
2003-05-01
影响因子：
5.8
通讯作者：
Gretzer, C
中科院分区：
文献类型：
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作者：
Tengvall, P;Jansson, E;Gretzer, C
通过乙基-二甲基-氨丙基碳二亚胺和N-羟基-琥珀酰亚胺偶联化学，由纤维蛋白原、白蛋白、IgG、纤维蛋白原和过氧化氢酶的组合以及硅上血浆制备交联的多层蛋白质膜。将4-70 nm厚的膜置于血浆中，并在孵育5或60分钟后通过零椭圆光度法测量额外的蛋白质沉积。补体系统的激活和凝血的内在途径通过随后抗C3c、抗C3d、抗备解素和抗HMWK在表面结合血浆顶部的结合来指示。分析单核细胞培养物中膜联蛋白 V、碘化丙啶和 4,6-二脒基-2-苯基吲哚阳性细胞的比例，以及肿瘤坏死因子 α (TNF-α) 和白细胞介素 10 (IL-10) 的分泌。结果表明，众所周知的蛋白质偶联技术可用于制备厚度控制良好的蛋白质层。这些层具有较低的血浆接触活化作用，并在单核细胞培养物中诱导不同程度的 TNF-α 和 IL-10 释放。 (C) 2002 Elsevier Science B.V. 保留所有权利。
Crosslinked multilayer protein films were prepared from fibrinogen, albumin, IgG, a combination of fibrinogen and catalase, and blood plasma on silicon by ethyl-dimethyl-aminopropylcarbodiimide and N-hydroxy-succinimide coupling chemistry. The 4-70 nm thick films were placed in blood plasma and the additional protein deposition measured by null ellipsometry after 5 or 60 min of incubation. The activation of the complement system and intrinsic pathway of coagulation were indicated through the subsequent binding of anti-C3c, anti-C3d, anti-properdin and anti-HMWK on top of the surface bound blood plasma. The proportion of Annexin V, Propidium Iodide and 4,6-diamidino-2-phenylindole positive cells, and the secretion of tumor necrosis factor alpha (TNF-alpha) and interleukin-10 (IL-10) were analysed in a monocyte culture. The results show that well known protein coupling techniques can be used for the preparation of protein layers with well controlled thickness. The layers possess low contact activation of blood plasma and induce different release of TNF-alpha and IL-10 in monocyte cultures. (C) 2002 Elsevier Science B.V. All rights reserved.