Special Section on Nanobio-Based Optical Sensing and Imaging

Site-directed immobilization of antibody using EDC-NHS-activated protein A on a bimetallic-based surface plasmon resonance chip

[+] Author Affiliations
Young-Soo Sohn

Catholic University of Daegu, Department of Biomedical Engineering, 13-13 Hayang-ro, Hayang-eup, Gyeongsan-si, Gyeongbuk 712-702, Republic of Korea

Yeon Kyung Lee

Catholic University of Daegu, Department of Biomedical Engineering, 13-13 Hayang-ro, Hayang-eup, Gyeongsan-si, Gyeongbuk 712-702, Republic of Korea

J. Biomed. Opt. 19(5), 051209 (Jan 13, 2014). doi:10.1117/1.JBO.19.5.051209
History: Received July 29, 2013; Revised November 25, 2013; Accepted December 13, 2013
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Abstract.  The characteristics of a waveguide-coupled bimetallic surface plasmon resonance (WcBiM SPR) sensor using (3-dimethylaminopropyl)-3-ethylcarbodiimide(EDC)-N-hydroxysuccinimide(NHS)-activated protein A was investigated, and the detection of IgG using the EDC-NHS-activated protein A was studied in comparison with protein A and a self-assembled monolayer (SAM). The WcBiM sensor, which has a narrower full width at half maximum (FWHM) and a steeper slope, was selected since it leads to a larger change in the reflectance in the intensity detection mode. A preparation of the EDC-NHS-activated protein A for site-directed immobilization of antibodies was relative easily compared to the engineered protein G and A. In antigen–antibody interactions, the response to IgG at the concentrations of 50, 100, and 150ng/ml was investigated. The results showed that the sensitivity of the WcBiM sensor using the EDC-NHS-activated protein A, protein A, and SAM was 0.0185 [%/(ng/ml)], 0.0065 [%/(ng/ml)], and 0.0101 [%/(ng/ml)], respectively. The lowest detectable concentrations of IgG with the EDC-NHS-activated protein A, protein A, and SAM were 4.27, 12.83, and 8.24ng/ml, respectively. Therefore, the increased sensitivity and lower detection capability of the WcBiM SPR chip with the EDC-NHS-activated protein A suggests that it could be used in early diagnosis where the trace level concentrations of biomolecules should be detected.

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© 2014 Society of Photo-Optical Instrumentation Engineers

Citation

Young-Soo Sohn and Yeon Kyung Lee
"Site-directed immobilization of antibody using EDC-NHS-activated protein A on a bimetallic-based surface plasmon resonance chip", J. Biomed. Opt. 19(5), 051209 (Jan 13, 2014). ; http://dx.doi.org/10.1117/1.JBO.19.5.051209


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