Preparation of a Universal Blood Donor Type

摘要

We had previously reported on the purification of the alpha(1 to 3)-N-acetyl-galactosaminidase from Cl. perfringens (8000 fold) and of the alpha(1 to 3)-galactosidases, B-zyme, from Cl. sporogenes (2500 fold). Both enzymes were, nonetheless demonstrated to be still impure by polyacrylamide gel electrophoresis. The purest preparation of the A-zyme was still contaminated with related glycosidases and they even co-migrated on polyacrylamide gel electrophoresis. This, together with other observations, implicated a multi-enzyme complex involving Sh reversible reaction S-S interactions. The presence of such interactions was demonstrated in the A-zyme and the other glycosidases, co-purifying with it, as well as with the B-zyme. With this knowledge, it was possible to exploit these properties to purify both the A-zyme and B-zyme. The applicability of immuno-affinity chromatography to the purification and separation of the exoglycosidases was also explored. A measure of success was attained. The development of this approach was arrested by the greater success we had by the above mentioned methods. The action of the exo-glycosidases; alpha(1 to 3)-D-galactosidase, alpha(1 to 3)-N-acetyl-D-galactosaminidase and alpha(1 to 2)L-fucosidase, on the blood group substances and oligosaccharides appear to be regulated by steric hindrance.