| 摘要 |
PURPOSE:To improve measuring accuracy of an enzyme immunity of hormone and to measure automatically, by reusing an insoluble carrier utilizing an exchange reaction of S-S bond and SH radical and adding a hydrophobic protein and salts to measuring system. CONSTITUTION:Tagged hormone combined enzyme such as beta-galactosidase with a hormone, insulin, etc. is prepared. On the one hand, glutathione-2-pyridylsulfide is incorporated into an insoluble carrier such as ''Cephadex G25 '' and a bead or tube is prepared making S-S bond of said carrier and antiimmunogloblin. Said tagged hormone is added to 10mM phosphoric acid buffer solution of 7pH incorporated, for example, 0.3M NaCl, 1mM MgCl2, 0.5% gelatine, 0.1% cattle blood serum albumin, 0.1% NaN3 and 2mM N-ethylmaleiimid to human blood serum to be measured and said solution is let flowed to said tubular carrier and then, enzyme is combined. Next, the S-S bond is exchanged for a S-H radical by a reducing agent and combined body of enzyme-antibody-hormone is eluted and then, hormone is determined by measuring the activity of enzyme in an eluate. Inactive antibody is reused by making S-S bond of the antibody again. |
