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J. Microbiol. Biotechnol. 2006; 16(1): 141-144

Published online January 28, 2006

Copyright © The Korean Society for Microbiology and Biotechnology.

Antibody Layer Fabrication for Protein Chip to Detect E. coli O157:H7, Using Microcontact Printing Technique

Kim, Hun-Soo , Young-Min Bae 1, Young-Kee Kim 2, Byung-Keun Oh 2 and Jeong-Woo Choi 3*

Department of Chemical and Biomolecular Engineering, and Interdisciplinary Program of Integrated Biotechnology, Sogang University, C.P.O. Box 1142, Seoul 100-611, Korea, Technology Planning Team, Corporate R&D Center, Samsung SDI Co. Ltd, Gyeonggi-d, 1Department of Chemical and Biomolecular Engineering, and Interdisciplinary Program of Integrated Biotechnology, Sogang University, C.P.O. Box 1142, Seoul 100-611, Korea, 2Department of Chemical Engineering, Hankyong National University, Kyonggi-Do 456-749, Korea, 3Department of Chemical and Biomolecular Engineering, and Interdisciplinary Program of Integrated Biotechnology, Sogang University, C.P.O. Box 1142, Seoul 100-611, Korea

Abstract

An antibody layer was fabricated to detect Escherichia coli O157:H7. The micropattern of 16-mercaptohexadecanoic acid (16-MHDA) as alkylthiolate was formed on the gold surface by using the PDMS stamp with microcontact printing $({\mu}CP)$ techniques. In order to form antibody patterns on the template, protein G was chemically bound to the 16-MHDA patterns, and antibody was adsorbed on a self-assembled protein G layer. The formation of the 16-MHDA micropattern, self-assembled protein G layer and antibody pattern on Au substrate was confirmed by surface plasmon resonance (SPR) spectroscopy. Finally, the micropatterning method was applied to fabricate the antibody probe for detection of E. coli O157:H7, and monitoring of antigen by using this probe was successfully achieved.

Keywords: Antibody probe, protein G, mercaptohexadecanoic acid, microcontact printing, micropattern, E. coli O157:H7