Protein array consisting of sol-gel bioactive platform for detection of E. coli O157:H7.
Sol-gel-derived bioactive platform was fabricated for detection of pathogenic microbes, E. coli O157:H7. Design flexibility of sol-gel technique and ease of fabrication can fulfill to create the surfaces with structural and chemical features that are compatible with biomaterials such as antibody, enzymes, etc. In this study, the bioactive platform was prepared based on the silica gels, which were produced by hydrolyzing tetraethylorthosilane (TEOS) in ethanol. The mercaptopropyl triethoxysilane (MPTS) was mixed with the TEOS solution for the surface functionalization of bioactive platform. During TEOS hydrolysis, the modified thin film was prepared by sol-gel dip coating. Antibody against E. coli O157:H7 was immobilized with a configuration of protein array using piezo-type dispensing system. Surface morphology of the prepared bioactive platform was analyzed using atomic force microscopy (AFM). The antibody-antigen interaction was investigated with fluorescence microscopy and sandwich type immunoassay using fluorescein isothiocyanate (FITC)-labeled antibody. The results showed that antibody was sequestered within the sol-gel-derived bio-gel due to physical adsorption. The measurement of E. coli O157:H7 was done using the fabricated antibody surface. The fluorescence intensity was proportional to the concentration of E. coli O157:H7, of which the detection limit was 10(2)CFU/ml.
Lee
Woochang
W
Interdisciplinary Program of Integrated Biotechnology, Sogang University, 1 Shinsu-Dong, Mapo-Gu, Seoul 121-742, Republic of Korea.
Park
Ki-Suk
KS
Kim
Yong-Wan
YW
Lee
Won Hong
WH
Choi
Jeong-Woo
JW
D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000737:chemistry* / Q000276:immunology; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D015169:Colony Count, Microbial / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D019453:Escherichia coli O157 / Q000276:immunology* / Q000302:isolation & purification*; D005782:Gels / Q000737:chemistry; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D044367:Phase Transition; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods; D013050:Spectrometry, Fluorescence / Q000295:instrumentation* / Q000379:methods
D000911:Antibodies, Monoclonal; D020099:Coated Materials, Biocompatible; D005782:Gels