Ultrasensitive immunosensing of tuberculosis CFP-10 based on SPR spectroscopy |
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Authors: | Seong Cheol HongAuthor Vitae Hongxia ChenAuthor Vitae Jaewook LeeAuthor VitaeHye-Kyung ParkAuthor Vitae Young Sik KimAuthor VitaeHyun-Chul ShinAuthor Vitae Cheol-Min KimAuthor VitaeTae Jung ParkAuthor Vitae Seok Jae LeeAuthor VitaeKwangnak KohAuthor Vitae Hwa-Jung KimAuthor VitaeChulhun L. ChangAuthor Vitae Jaebeom LeeAuthor Vitae |
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Affiliation: | a Department of Nanomedical Engineering, Pusan National University, Miryang 627-706, Republic of Korea b School of Life Sciences, Shanghai University, Shanghai 200444, China c School of Medicine, Pusan National University, Yangsan 626-770, Republic of Korea d Department of Chemistry Education, Korea National University of Education, Cheongwon 363-791, Republic of Korea e Bioprocess Engineering Research Center, Center for Systems & Synthetic Biotechnology, and Institute for the BioCentury, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea f NEMS-Bio Team, National NanoFab Center, 335 Gwahangno, Yuseong-gu, Daejeon 305-806, Republic of Korea g Department of Microbiology and Research Institute for Medical Science, College of Medicine, Chungnam National University, Daejeon 301-747, Republic of Korea |
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Abstract: | An antigen (Ag), CFP-10, found in tissue fluids of tuberculosis (TB) patients may be an ultimate candidate for use as a sensitive TB marker with a sensing method for early simplified diagnosis of TB. In this study, chemical and optical optimizations were carried out using novel immuno-materials for establishment of a self-assembled surface plasmon resonance (SPR) optical immunosensor system for detection of CFP-10, which is valuable for pre-clinical work, prior to conduct of massive clinical observations. For creation of a simple sensing interface, a monoclonal antibody (anti-CFP-10) was immobilized directly on a gold surface, followed by blocking with cystamine. Orientation and accessibility of anti-CFP-10 were assessed by the selective binding of CFP-10. Recent results indicate that the reusability of the sensor chip adopting the cystamine method was found to be preferable to other immobilization methods. A linear relationship was well correlated between SPR angle shift and CFP concentrations in the range from 100 ng mL−1 to 1 μg mL−1. Modification of the SPR chip with antibody provides a simple experimental platform for investigation of isolated proteins under experimental conditions resembling those of their native environment. |
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Keywords: | Tuberculosis (TB) CFP-10 Surface plasmon resonance (SPR) spectroscopy Biosensor |
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