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KMID : 1094720080130040445
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Biotechnology and Bioprocess Engineering
2008 Volume.13 No. 4 p.445 ~ p.452
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Selective removal of anti-α-Gal antibodies from human serum by using synthetic α-Gal epitope on a core-shell type resin
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Jang Kyoung-Soon
Chung Woo-Jae
Kim Hyun-Ki
Kim Yun-Gon
Lee Yoon-Sik
Kim Byung-Gee
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Abstract
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A novel α-Gal resin was chemo-enzymatically synthesized for the efficient adsorption of anti-α-Gal antibodies in human serum for xenotransplantation. To covalently conjugate a hexanoate linker with lactose and N-acetylglucosamine, both acceptor sugars were acetylated and brominated. Then, α-and β-galactoses were sequentially added to the linker-containing saccharides at their non-reducing ends by using recombinant α-(1,3)-and β-(1,4)-galactosyltransferases from E. coli. Finally, the synthesized α-Gal derivatives were immobilized on HiCore, a core-shell type resin, that was functionalized with amino groups on the shell region, as a packing material on-column. Using this method we were able to demonstrate that the α-Gal HiCore resin had a reduced level of non-specific protein adsorption compared with the commercially available polystyrene supports, TentaGel, and agarose-based supports, when Lectin BS-I was used as the model binding protein. Furthermore, the α-Gal HiCore resin was more efficient at eliminating anti-α-Gal IgGs from the total human IgGs through immunoadsorption than the other two α-Gal resins, α-Gal TentaGel and α-Gal agarose. The α-Gal HiCore resin developed in this study can be utilized in a wide range of applications including ex vivo immunoadsorption and as a quantitative assay of anti-Gal antibody in human sera.
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KEYWORD
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α-Gal epitope, core-shell type resin, HiCore, anti-α-Gal antibody, hyperacute rejection, xenotransplantation, immunoadsorption
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