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Surface Plasmon Resonance Immunoassay for Biotin Determination on a Home-made Instrument
Available online at www.sciencedirect.com
ScienceDirect Procedia Technology 27 (2017) 87 – 88
Biosensors 2016
Surface plasmon resonance immunoassay for biotin determination on a home-made instrument Xing Chen*, Lulu Zhang, Dafu Cui State Key Laboratory of Transducer Tech., Institute of Electronics, Chinese Academy of Sciences, Beijing, 100190, China
Abstract In this paper, a home-made SPR analytical system and a denaturalized bovine serum albumin (dBSA) SPR chip were used to detect biotin based on the method of an inhibition immunoassay. The experimental was compared with a commercial protocol. The conditions of regeneration were optimized. The dBSA SPR chip was fabricated by a self-assembling method and the derivative of biotin was immobilized on the dBSA SPR chip by using a on-line mode. A commercial instrument and a dextranSPR-chip (CM5) were used to implement the same experiments. Based on the experiments of immune reactions between biotin antibody and biotin derivative, the biotin antibody concentration was optimized at about 5µg/mL in order to obtain highly sensitivity, which was used in the followed inhibition immunoassays. The lowest detection limit for biotin is 0.1µg/mL with a detection range of 0.1-1000 µg/ml by using our home-made instrument and the dBSA-SPR-chip. The detection results of our instrument and dBSA SPR chip are comparable with the commercial protocol. Our home-made instrument and dBSA-SPR-chips can be also applied to detect other small molecules based on the indirect inhibition immunoassay. © by Elsevier Ltd. by This is an open The Authors. Published Elsevier Ltd.access article under the CC BY-NC-ND license © 2017 2016Published (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the organizing committee of Biosensors 2016. Peer-review under responsibility of the organizing committee of Biosensors 2016 Keywords: Surface plasmon resonance(SPR); inhibition immunoassay; small molecule detection
2212-0173 © 2017 Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the organizing committee of Biosensors 2016 doi:10.1016/j.protcy.2017.04.037
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Xing Chen et al. / Procedia Technology 27 (2017) 87 – 88
(a)
(b)
Fig. 1. Biotin assay standard curve with a detection range of 0.1ng/ml-1000 µg/ml (a) and with a detection range of 100ng/ml-1000 µg/ml (b) by using our home-made instrument and the dBSA-SPR-chip.
Acknowledgements The authors greatly acknowledge the financial support from National High-tech R&D Program of China (2014 AA 022303). This work is also sponsored by the Major National Scientific Research Plan under Grant number (2014CB744600) and National Science Foundation of China under Grant number 61571420, 31571007 and 81371711. References [1] Chen X, Zhang LL, Cui DF. Surface plasmon resonance immunoassay for cortisol determination with a self-assembling denaturalised bovine serum albumin layer on surface plasmon resonance chip, Micro & Nano Letters, 2016, 11(1): 20-23 [2] Cui DF, Cai HY, Wang JB, et al., Chinese Patent Appl. No. CN 200610066542.4, 2006
ScienceDirect Procedia Technology 27 (2017) 87 – 88
Biosensors 2016
Surface plasmon resonance immunoassay for biotin determination on a home-made instrument Xing Chen*, Lulu Zhang, Dafu Cui State Key Laboratory of Transducer Tech., Institute of Electronics, Chinese Academy of Sciences, Beijing, 100190, China
Abstract In this paper, a home-made SPR analytical system and a denaturalized bovine serum albumin (dBSA) SPR chip were used to detect biotin based on the method of an inhibition immunoassay. The experimental was compared with a commercial protocol. The conditions of regeneration were optimized. The dBSA SPR chip was fabricated by a self-assembling method and the derivative of biotin was immobilized on the dBSA SPR chip by using a on-line mode. A commercial instrument and a dextranSPR-chip (CM5) were used to implement the same experiments. Based on the experiments of immune reactions between biotin antibody and biotin derivative, the biotin antibody concentration was optimized at about 5µg/mL in order to obtain highly sensitivity, which was used in the followed inhibition immunoassays. The lowest detection limit for biotin is 0.1µg/mL with a detection range of 0.1-1000 µg/ml by using our home-made instrument and the dBSA-SPR-chip. The detection results of our instrument and dBSA SPR chip are comparable with the commercial protocol. Our home-made instrument and dBSA-SPR-chips can be also applied to detect other small molecules based on the indirect inhibition immunoassay. © by Elsevier Ltd. by This is an open The Authors. Published Elsevier Ltd.access article under the CC BY-NC-ND license © 2017 2016Published (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the organizing committee of Biosensors 2016. Peer-review under responsibility of the organizing committee of Biosensors 2016 Keywords: Surface plasmon resonance(SPR); inhibition immunoassay; small molecule detection
2212-0173 © 2017 Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the organizing committee of Biosensors 2016 doi:10.1016/j.protcy.2017.04.037
88
Xing Chen et al. / Procedia Technology 27 (2017) 87 – 88
(a)
(b)
Fig. 1. Biotin assay standard curve with a detection range of 0.1ng/ml-1000 µg/ml (a) and with a detection range of 100ng/ml-1000 µg/ml (b) by using our home-made instrument and the dBSA-SPR-chip.
Acknowledgements The authors greatly acknowledge the financial support from National High-tech R&D Program of China (2014 AA 022303). This work is also sponsored by the Major National Scientific Research Plan under Grant number (2014CB744600) and National Science Foundation of China under Grant number 61571420, 31571007 and 81371711. References [1] Chen X, Zhang LL, Cui DF. Surface plasmon resonance immunoassay for cortisol determination with a self-assembling denaturalised bovine serum albumin layer on surface plasmon resonance chip, Micro & Nano Letters, 2016, 11(1): 20-23 [2] Cui DF, Cai HY, Wang JB, et al., Chinese Patent Appl. No. CN 200610066542.4, 2006