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Acta Physico-Chimica Sinica  2010, Vol. 26 Issue (10): 2821-2827    DOI: 10.3866/PKU.WHXB20101016
BIOPHYSICAL CHEMISTRY     
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. 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
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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 wordsProtein microarray      Antibody immobilization      Sandwich reaction      Enzyme-linked immunosorbent assay      Amino surface      Glutaraldehyde      Poly-L-lysine     
Received: 22 April 2010      Published: 27 September 2010
MSC2000:  O647  
Fund:  

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).

Corresponding Authors: GAO Yun-Hua, XU Bai     E-mail: yhgao@mail.ipc.ac.cn, nanobioic@gmail.com
Cite this article:

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. Acta Physico-Chimica Sinica, 2010, 26(10): 2821-2827.

URL:

http://www.whxb.pku.edu.cn/Jwk_wk/wlhx/10.3866/PKU.WHXB20101016     OR     http://www.whxb.pku.edu.cn/Jwk_wk/wlhx/Y2010/V26/I10/2821

1. Zhu, H.; Snyder, M. Curr. Opin. Chem. Biol., 2001, 5: 40
2. MacBeath, G. Nat Genet, 2002, 32(suppl.): 526
3. Templin, M. F.; Stoll, D.; Schrenk, M.; Traub, P. C.; Võhringer, C. F.; Joos, T. O. Trends Biotechnol., 2002, 20: 160
4. Ng, J. H.; Ilag, L. L. Journal of Cellular and Molecular Medicine, 2002, 6: 329
5. Wilson, D. S.; Nock, S. Angew. Chem. Int. Edit., 2003, 42: 494
6. Angenendt, P. Drug Discovery Today, 2005, 10: 503
7. Kusnezow, W.; Hoheisel, J. D. Biotechniques, 2002, 33(suppl.): 14
8. Liotta, L. A.; Espina, V.; Mehta, A. I.; Calvert, V.; Rosenblatt, K.; Geho, D.; Munson, P. J.; Young, L.; Wulfkuhle, J.; Petricoin III, E. F. Cancer Cell, 2003, 3: 317
9. Lutanie, E.; Voegel, J. C.; Schaaf, P.; Freund, M.; Cazenave, J. P.; Schmitt, A. Proc. Natl. Acad. Sci. U. S. A., 1992, 89: 9890
10. Vijayendran, R. A.; Leckband, D. E. Anal. Chem., 2000, 73: 471
11. Rossier, J. S.; Gokulrangan, G.; Girault, H. H.; Svojanovsky, S.; Wilson, G. S. Langmuir, 2000, 16: 8489
12. Angenendt, P.; Glõkler, J.; Murphy, D.; Lehrach, H.; Cahill, D. J. Anal. Biochem., 2002, 309: 253
13. Kusnezow, W.; Hoheisel, J. D. J. Mol. Recognit., 2003, 16: 165
14. Seong, S. Y. Clin Diagn Lab Immunol, 2002, 9: 927
15. Kusnezow, W.; Jacob, A.; Walijew, A.; Diehl, F.; Hoheisel, J. D. Proteomics, 2003, 3: 254
16. Niemeyer, C. M.;Wacker, R.; Adler, M. Nucl. Acids Res., 2003, 31: e90
17. Peluso, P.;Wilson, D. S.; Do, D.; Tran, H.; Venkatasubbaiah, M.; Quincy, D.; Heidecker, B.; Poindexter, K.; Tolani, N.; Phelan, M.; Witte, K.; Jung, L. S.;Wagner, P.; Nock, S. Anal. Biochem., 2003, 312: 113
18. Ramachandran, N.; Hainsworth, E.; Bhullar, B.; Eisenstein, S.; Rosen, B.; Lau, A. Y.;Walter, J. C.; LaBaer, J. Science, 2004, 305: 86
19. Oh, S. J.; Hong, B. J.; Choi, K. Y.; Park, J. W. OMICS, 2006, 10: 327
20. Iwata, R.; Satoh, R.; Iwasaki, Y.; Akiyoshi, K. Colloids Surf. B, 2008, 62: 288
21. Hermanson, G. T. Bioconjugate techniques. 2nd ed. Rockford, Illinois: Academic Press, 2008
22. Haab, B. B.; Dunham, M. J.; Brown, P. O. Genome Biol., 2001, 2: 0004.1
23. Stillman, B. A.; Tonkinson, J. L. Biotechniques, 2000, 29: 630
24. Qian,W.; Yao, D.; Yu, F.; Xu, B.; Zhou, R.; Bao, X.; Lu, Z. Clin. Chem., 2000, 46: 1456
25. MacBeath, G.; Schreiber, S. L. Science, 2000, 289: 1760
26. Piehler, J.; Brecht, A.; Geckeler, K. E.; Gauglitz, G. Biosens. Bioelectron., 1996, 11: 579
27. Penzol, G.; Armisén, P.; Fernández-Lafuente, R.; Rodés, L.; Guisán, J. M. Biotechnol. Bioeng., 1998, 60: 518
28. Afanassiev, V.; Hanemann, V.;Wolfl, S. Nucl. Acids Res., 2000, 28: e66
29. Piletsky, S. A.; Matuschewski, H.; Schedler, U.; Wilpert, A.; Piletska, E. V.; Thiele, T. A.; Ulbricht, M. Macromolecules, 2000, 33: 3092
30. Wilson, D. S.; Nock, S. Curr. Opin. Chem. Biol., 2002, 6: 81
31. Neubert, H.; Jacoby, E. S.; Bansal, S. S.; Iles, R. K.; Cowan, D. A.; Kicman, A. T. Anal. Chem., 2002, 74: 3677
32. Gupta, B.; Plummer, C.; Bisson, I.; Frey, P.; Hilborn, J. Biomaterials, 2002, 23: 863
33. Stine, R.; Cole, C. L.; Ainslie, K. M.; Mulvaney, S. P.;Whitman, L. J. Langmuir, 2007, 23: 4400
34. Gibson, T. D.;Woodward, J. R. Protein stabilization in biosensor systems//Biosensors and chemical sensors.Washington, DC: American Chemical Society, 1992: 40
35. Mateo, C.; Torres, R.; Fernández-Lorente, G.; Ortiz, C.; Fuentes, M.; Hidalgo, A.; López-Gallego, F.; Abian, O.; Palomo, J. M.; Betancor, L.; Pessela, B. C. C.; Guisan, J. M.; Fernández-Lafuente, R. Biomacromolecules, 2003, 4: 772
36. Lee, S. D.; Hsiue, G. H.; Chang, P. C. T.; Kao, C. Y. Biomaterials, 1996, 17: 1599
37. Kato, K.; Ikada, Y. Biotechnol. Bioeng., 1995, 47: 557
38. Lee,W.; Oh, B. K.; Bae, Y. M.; Paek, S. H.; Lee, W. H.; Choi, J. W. Biosensors and Bioelectronics, 2003, 19: 185
39. Turková, J. J. Chromatogr. B, 1999, 722: 11
40. Jung, Y.; Lee, J. M.; Jung, H.; Chung, B. H. Anal. Chem., 2007, 79: 6534
41. Võlimaa, L. Streptavidin———a versatile binding protein for solid- phase immunoassays [D]. Turku: University of Turku, 2008
42. Lue, R. Y. P.; Chen, G. Y. J.; Zhu, Q.; Lesaicherre, M. -L.; Yao, S. Q. Site-specific immobilization of biotinylated proteins for protein microarray analysis//Protein arrays. Totowa: Humana Press Inc., 2004: 85

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