naa a22 4500 510820360 CHVBK 20180411083528.0 cr unu---uuuuu 180411e20131001xx s 000 0 eng 10.1007/s11814-013-0117-5 doi (NATIONALLICENCE)springer-10.1007/s11814-013-0117-5 A comparative study on antibody immobilization strategies onto solid surface [Elektronische Daten] [Ji Lee, Jeong Seo, Chang Kim, Yunkyeoung Kwon, Jeong Ha, Suk Choi, Hyung Cha] Antibody immobilization onto solid surface has been studied extensively for a number of applications including immunoassays, biosensors, and affinity chromatography. For most applications, a critical consideration regarding immobilization of antibody is orientation of its antigen-binding site with respect to the surface. We compared two oriented antibody immobilization strategies which utilize thiolated-protein A/G and thiolated-secondary antibody as linker molecules with the case of direct surface immobilization of thiol-conjugated target antibody. Antibody immobilization degree and surface topography were evaluated by surface plasmon resonance and atomic force microscope, respectively. Protein A/G-mediated immobilization strategy showed the best result and secondary antibody-mediated immobilization was the worst for the total immobilization levels of target antibodies. However, when considering real-to-ideal ratio for antigen binding, total target antigen binding levels (oriented target antibody immobilization levels) had the following order: secondary antibody-mediated immobilization>protein A/G-mediated immobilization>direct thiol-conjugated immobilization. Thus, we confirmed that protein A/G- and secondary antibody-mediated strategies, which consider orientation of target antibody immobilization, showed significantly high antigen binding efficiencies compared to direct random immobilization method. Collectively, the oriented antibody immobilization methods using linker materials could be useful in diverse antibody-antigen interaction-involved application fields. Korean Institute of Chemical Engineers, Seoul, Korea, 2013 Antibody Immobilization nationallicence Orientation nationallicence Thiolation nationallicence Protein A/G nationallicence Secondary Antibody nationallicence Surface Plasmon Resonance nationallicence Atomic Force Microscopy nationallicence Lee Ji Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut Seo Jeong Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut Kim Chang Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut Kwon Yunkyeoung Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut Ha Jeong School of Environmental Science and Technology, Pohang University of Science and Technology, 790-784, Pohang, Korea aut Choi Suk Department of Biological and Environmental Engineering, Semyung University, 390-711, Jecheon, Korea aut Cha Hyung Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut Korean Journal of Chemical Engineering Springer US; http://www.springer-ny.com 30/10(2013-10-01), 1934-1938 0256-1115 30:10<1934 2013 30 11814 https://doi.org/10.1007/s11814-013-0117-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11814-013-0117-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lee Ji Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut NATIONALLICENCE 700 1- Seo Jeong Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut NATIONALLICENCE 700 1- Kim Chang Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut NATIONALLICENCE 700 1- Kwon Yunkyeoung Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut NATIONALLICENCE 700 1- Ha Jeong School of Environmental Science and Technology, Pohang University of Science and Technology, 790-784, Pohang, Korea aut NATIONALLICENCE 700 1- Choi Suk Department of Biological and Environmental Engineering, Semyung University, 390-711, Jecheon, Korea aut NATIONALLICENCE 700 1- Cha Hyung Department of Chemical Engineering, Pohang University of Science and Technology, 790-784, Pohang, Korea aut NATIONALLICENCE 773 0- Korean Journal of Chemical Engineering Springer US; http://www.springer-ny.com 30/10(2013-10-01), 1934-1938 0256-1115 30:10<1934 2013 30 11814 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer