硅基芯片表面化学性质对蛋白质固定化的影响

引用本文: 周稳稳, 廉洁, 胡科家, 高云华, 徐百. 硅基芯片表面化学性质对蛋白质固定化的影响[J]. 物理化学学报, 2010, 26(10): 2821-2827. doi: 10.3866/PKU.WHXB20101016 shu

Citation: ZHOU Wen-Wen, LIAN Jie, HU Ke-Jia, GAO Yun-Hua, XU Bai. Effect of Surface Chemical Properties of a Silicon Chip on Antibody Immobilization[J]. Acta Physico-Chimica Sinica, 2010, 26(10): 2821-2827. doi: 10.3866/PKU.WHXB20101016 shu

硅基芯片表面化学性质对蛋白质固定化的影响

基金项目:
国家自然科学基金(20975106) (20975106)

中国科学院知识创新工程重要方向项目(KJCX2-YW-M15) 及国家重大专项(2008ZX08012-001) 资助 (KJCX2-YW-M15) 及国家重大专项(2008ZX08012-001)

摘要:

制备蛋白质芯片的关键在于将蛋白质固定到芯片表面并保持其生物学活性. 本实验中, 我们分别采用物理吸附、直接化学固定、加入间隔臂化学固定和生物亲和作用固定的方法将癌胚抗原(CEA)抗体固定到硅基芯片的二氧化硅表面. 基于抗原-抗体的特异性相互作用, 利用双抗体夹心酶联免疫法(ELISA)评价各种方法固定抗体的效果. 实验结果表明, 在修饰有氨基的表面采用戊二醛作为偶联试剂固定CEA 抗体具有最高的偶联效率, 引入多聚赖氨酸(poly-L-lysine)作为间隔臂可以显著增强固定效果, 并可进一步降低非特异性吸附. 而利用生物亲和作用固定CEA抗体也可获得较好的固定效果,但是非特异性吸附较严重.

关键词:

English

Effect of Surface Chemical Properties of a Silicon Chip on Antibody Immobilization

1.
1. Key Laboratory of Photochemical Conversion and Optoelectronic Materials, Technical Institute of Physics and Chemistry,Chinese Academy of Sciences, Beijing 100190, P. R. China;
2. Graduate University of Chinese Academy of Sciences,Beijing 100049, P. R. China
Received Date: 22 April 2010
Available Online: 27 September 2010
Fund Project:
国家自然科学基金(20975106)

中国科学院知识创新工程重要方向项目(KJCX2-YW-M15) 及国家重大专项(2008ZX08012-001) 资助

Abstract:

The key to constructing a protein microarray is the stable immobilization of proteins and the retention of their biological activities. In this study, immobilization of the carcinoembryonic antigen (CEA) antibody onto a silicon dioxide surface was investigated by physical adsorption, direct chemical covalent conjugation, spacer-added chemical covalent conjugation, and biological affinity interactions. Based on the specific antibody-antigen interactions, the sandwich reaction, enzyme-linked immunosorbent assay (ELISA) was chosen to evaluate various immobilization strategies. The most efficient immobilization strategy was with glutaraldehyde as a coupling reagent between the CEA antibody and the amino surface. The attachment of a spacer-armcomprising poly-L-lysine significantly improved the immobilization efficiency and simultaneously decreased nonspecific adsorption. High immobilization efficiency and stronger nonspecific adsorption were also observed when the CEA antibody was immobilized by bioaffinity interactions.

Key words:

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

硅基芯片表面化学性质对蛋白质固定化的影响

English

Effect of Surface Chemical Properties of a Silicon Chip on Antibody Immobilization

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

硅基芯片表面化学性质对蛋白质固定化的影响

English

Effect of Surface Chemical Properties of a Silicon Chip on Antibody Immobilization

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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中国化学会秘书处

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