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

Abstract

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 k-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 bK=Åçäá. Finally, the synthesized α-Gal derivatives were immobilized on eáCore, 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 eáCore 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 eáCore 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 eáCore resin developed in this study can be utilized in a wide range of applications including Éñ=îáîç immunoadsorption and as a quantitative assay of anti-Gal antibody in human sera. © KSBB hÉóïçêÇëW=αJd~ä=ÉéáíçéÉI=ÅçêÉJëÜÉää=íóéÉ=êÉëáåI=Hi`çêÉI=~åíáJαJd~ä=~åíáÄçÇóI=ÜóéÉê~ÅìíÉ=êÉàÉÅíáçåI=ñÉåçíê~åëéä~åí~íáçåI= = áããìåç~Çëçêéíáçå= = = = =

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2007–2008

“…For preparation of resin to remove anti-α-Gal antibodies from human serum (Jang, et al, 2008) C-Galactosides TOF (DHB CHCA)…”

Immobilization of UDP‐Galactose on an Amphiphilic Resin

“…Most sugars are attached to solid‐phase supports or resins through C1 with no nucleotide present . Sugars are only coupled to resins through C6 when it is used as a synthetic aid and are detached before the end of the synthesis .…”