Electrochemically oriented immobilization of antibody on poly-(2-cyano-ethylpyrrole)-coated gold electrode using a cyclic voltammetry |
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Authors: | Um Hyun-Ju Kim Mina Lee Sang-Hee Min Jiho Kim Hongik Choi Young-Woo Kim Yang-Hoon |
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Affiliation: | a Department of Microbiology, Chungbuk National University, 410 Sungbong-Ro, Heungduk-Gu, Cheongju 361-763, South Korea b Department of Chemical Engineering, Chonbuk National University, 664-14, 1-Ga, Duckjin-Dong, Duckjin-Gu, Jeonju 561-156, South Korea c R&D Division, Vitabio, Inc., 483 Expo-Ro, Yuseong-Gu, Daejeon 300-824, South Korea d Fuel Cell Research Center, Korea Institute of Energy Research, 71-2 Jang-dong, Yuseong-Ku, Daejeon 305-343, South Korea |
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Abstract: | Using quartz crystal microbalance (QCM) as an immunosensor, this work investigates the contribution of a cyclic voltammetry (CV) on the proper immobilization of antibodies with the aim of enhancing its target recognition and binding ability. Primarily, CV in the range of −0.1 to 0.9 V was applied to form a layer of poly-(2-cyano-ethylpyrrole) (PCEPy) on gold quartz crystal electrode. Then the efficiencies of antibodies (anti-IgG, AIgG) immobilized electrochemically with CV applied in 0-0.65 V were compared to those immobilized via physical adsorption, by observing relative affinity towards AIgG-Fab and AIgG-Fc fragments. The results showed antibody-AIgG-Fab interaction could be enhanced about 4 times when CV is applied (11.2 ± 1.3 vs 41.6 ± 3.4 relative fluorescence unit). On the contrary, physisorbed antibodies showed a higher degree of affinity towards AIgG-Fc indicating inappropriate orientations of physisorbed antibodies. AIgG immobilized PCEPy-gold QC electrode was characterized further for its sensitivity towards a new target bovine albumin with both a QCM and fluorescence measurement. Such electrode exhibited a good sensitivity as well as a large linear dynamic range, from 0.4 μg/ml to 1.0 μg/ml and from 0.5 μg/ml to 10.0 μg/ml, at QCM and fluorescence measurement, respectively. |
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Keywords: | Electrochemically oriented antibody immobilization Cyclic voltammetry Quartz crystal microbalance Antibody orientation Poly-(2-cyano-ethylpyrrole) |
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