Acta Phys. -Chim. Sin. ›› 2010, Vol. 26 ›› Issue (10): 2821-2827.doi: 10.3866/PKU.WHXB20101016

• BIOPHYSICAL CHEMISTRY • Previous Articles     Next Articles

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

ZHOU Wen-Wen1,2, LIAN Jie1,2, HU Ke-Jia1,2, GAO Yun-Hua1, XU Bai1   

  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:2010-04-22 Revised:2010-07-09 Published:2010-09-27
  • Contact: GAO Yun-Hua, XU Bai E-mail:yhgao@mail.ipc.ac.cn, nanobioic@gmail.com
  • Supported by:

    The project was supported by the National Natural Science Foundation of China (20975106), Funds of the Chinese Academy of Sciences for Key Topics in Innovation Engineering (KJCX2-YW-M15) and Important National Science & Technology Specific Projects, China (2008ZX08012-001).

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: Protein microarray, Antibody immobilization, Sandwich reaction, Enzyme-linked immunosorbent assay, Amino surface, Glutaraldehyde, Poly-L-lysine

MSC2000: 

  • O647