Oriented immobilization of antibodies with GST-fused multiple Fc-specific B-domains on a gold surface

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Oriented immobilization of antibodies with GST-fused multiple Fc-specific B-domains on a gold surface
Tai-Hwan Ha; S O Jung; Jeong Min Lee; Kun Yeong Lee; Y Lee; J S Park; Bong Hyun Chung
Bibliographic Citation
Analytical Chemistry, vol. 79, no. 2, pp. 546-556
Publication Year
We have constructed a novel platform for the oriented buildup of immunoglobulins on a gold surface for a surface plasmon resonance imaging microarray. To this end, genetically engineered glutathione S-transferase proteins bearing one, two, and three Fc-specific B-domains in protein G from Streptococci (GST-GB1, -GB2, and -GB3, respectively) were produced. In order to tether these GST-GBx proteins specifically, a novel glutathione-derivatized ligand (LA-GSH) was also synthesized from a biaminated tri(ethylene glycol) backbone. Each end of the backbone was further functionalized with a maleimide group for a glutathione modification and a lipoic acid for surface immobilization. The glutathione ligand demonstrated a negligible nonspecific protein adsorption toward other spectator proteins while showing a strong specific association toward GST-GBx proteins. This Fc-specific surface exhibited at least a 2-fold enhancement in the immunoglobulin density (from human and mouse) with its antigen capture capability totally conserved compared to a covalentiy tethered GBx proteins. A single antibody tethered on the GST-GB 3 is estimated to capture two antigens (enhanced green fluorescent protein), and this antigen capture ratio seems to be the most efficient value ever observed.
Amer Chem Soc
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Division of Bio Technology Innovation > Core Research Facility & Analysis Center > 1. Journal Articles
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