The development of protein chip using protein G for the simultaneous detection of various pathogens.
A protein chip using protein G for the simultaneous detection of various pathogens such as Escherichia coli O157:H7, Salmonella typhimurium, Yersinia enterocolitica, and Legionella pneumophila was developed. In order to endow the orientation of antibody molecules on solid surface, protein G was introduced. The protein G on gold (Au) surface modified with 11-mercaptoundecanoic acid (MUA) was arrayed and then four different kinds of monoclonal antibodies (Mabs) against pathogens (E. coli O157:H7, S. typhimurium, Y. enterocolitica, and L. pneumophila) on protein G spots were selectively arrayed using a microarrayer, and its spatial density was over 2400 spots cm(2). Using the constructed protein chip, the various pathogens such as E. coli O157:H7, S. typhimurium, Y. enterocolitica, and L. pneumophila could be detected by a sandwich method and its lowest detection limit for E. coli O157:H7 was 10(2) CFU/ml. The proposed fabrication technique of protein chip for the detection of various pathogens could be applied to construct other protein chips with a high efficiency.
Choi
Jeong-Woo
JW
Department of Chemical and Biomolecular Engineering, Sogang University, 1 Shinsu-dong, Mapo-gu, Seoul 121-742, Republic of Korea.
Kim
Young-Kee
YK
Oh
Byung-Keun
BK
D000907:Antibodies, Bacterial / Q000737:chemistry / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D015169:Colony Count, Microbial; D006046:Gold / Q000737:chemistry; D006090:Gram-Negative Bacteria / Q000145:classification / Q000276:immunology*; D016905:Gram-Negative Bacterial Infections / Q000382:microbiology*; D006801:Humans; D007118:Immunoassay; D018625:Microscopy, Atomic Force; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; C029152:G-substrate; D009419:Nerve Tissue Proteins; D006046:Gold