Preparation of a glycoprotein fraction from pooled human plasma and its evaluation as a substrate for the assay of Clostridium welchii (C. perfringens) neuraminidase.
A glycoprotein fraction (fraction VII) suitable for use as a substrate in assays of microbial neuraminidase was prepared from pooled human plasma. It is pasteurised during preparation to eliminate the risk of transmission of serum hepatitis. This results in polymerisation of some of the gamma1-acid glycoprotein, but fraction VII is shown to be an excellent substrate for the neuraminidase of Clostridium welchii (C. perfringens). A sensitive assay procedure is described. A number of factors may interfere with the measurement of NANA released by the action of microbial neuraminidase and procedures are described for evaluation of this problem. Fraction VII is easy to prepare, cheap and available in standard form in large amounts (inquiries should be addressed to J. K. S.); it is recommended for routine use as a convenient substrate for neuraminidase assays.
D003016:Clostridium perfringens / Q000201:enzymology*; D003470:Culture Media; D003956:Dialysis; D004305:Dose-Response Relationship, Drug; D006023:Glycoproteins / Q000032:analysis / Q000097:blood* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009439:Neuraminidase / Q000032:analysis* / Q000378:metabolism; D012794:Sialic Acids / Q000378:metabolism
D003470:Culture Media; D006023:Glycoproteins; D012794:Sialic Acids; D009439:Neuraminidase