Selective removal of anti-α-Gal antibodies from human serum by using synthetic α-Gal epitope on a core-shell type resin

• Published in: Biotechnology and bioprocess engineering Vol. 13; no. 4; pp. 445 - 452
• Main Authors: Jang, K.S. (Seoul National University, Seoul, Republic of Korea), Chung, W.J. (Seoul National University, Seoul, Republic of Korea), Kim, H.K. (Seoul National University, Seoul, Republic of Korea), Kim, Y.G. (Seoul National University, Seoul, Republic of Korea), Lee, Y.S. (Seoul National University, Seoul, Republic of Korea), Kim, B.G. (Seoul National University, Seoul, Republic of Korea), E-mail: byungkim@snu.ac.kr
• Format: Journal Article
• Language: English
• Published: Heidelberg The Korean Society for Biotechnology and Bioengineering 01.08.2008; 한국생물공학회
• Subjects: alpha-Gal epitope; anti-alpha-Gal antibody; Biotechnology; Chemistry; Chemistry and Materials Science; core-shell type resin; Escherichia coli; HiCore; hyperacute rejection; immunoadsorption; Industrial and Production Engineering; PROTEINAS; PROTEINE; PROTEINS; xenotransplantation; 생물공학; Hi
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-008-0141-1

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.