Quantitative assay of hepatitis B surface antigen by using surface plasmon resonance biosensor |
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Authors: | Sang Yoon Hwang Chang Hoon Yoon Jun Yeoung Jeon Sung Chul Choi Eun Kyu Lee |
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Affiliation: | (1) Bioprocessing Research Laboratory, Department of Chemical Engineering, Hanyang University, 426-791 Ansan, Korea;(2) Isu Chemicals, Inc., 137-714 Seoul, Korea;(3) DI Biotech, Ltd., 135-811 Seoul, Korea;(4) GE Team, GreenCross Vaccine Corp., 449-903 Yongln, Korea |
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Abstract: | We performed a basic experiment for the rapid, on-line, real-time measurement of hepatitis B surface antigen using a surface plasmon resonance biosensor. We immobilized anti-HBsAg (hepatitis B surface antigen) polyclonal antibody, as a ligand, to the dextran layer on a CM5 chip surface that had previously been activated byN-hydroxysuccinimide. A sample solution containing HBsAg was fed through a microfluidic channel, and the reflecting angle change due to the mass increase from the binding was detected. The binding characteristics between HBsAg and its polyclonal antibody followed the typical monolayer adsorption isotherm. When the entire immobilized antibody had interacted, no additional, non-specific binding occurred, suggesting the immunoreaction was very specific. The bound antigen per unit mass of the antibody was independent of the immobilized ligand density. No significant steric hindrance was observed at an immobilization density of approximately 17.6 ng/mm2. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the ligand was linear up to ca. 40 μg/mL. This linearity was much higher than that of the ELISA method. It appeared the antigen-antibody binding increased as the immobilized ligand density increased. In summary, this study showed the potential of this SPR biosensor-based method as a rapid, simple and multi-sample on-line assay. Once properly validated, it may serve as a more efficient method for HBsAg quantification for replacing the ELISA. |
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Keywords: | SPR biosensor HBsAg binding assay |
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