ENZYME-LINKED SANDWICH IMMUNOASSAY OF ORNITHINE DELTA-AMINOTRANSFERASE FROM RAT-LIVER USING ANTIBODY-COUPLED GLASS RODS AS SOLID-PHASE

被引:10
作者
HAMAGUCHI, Y
KATO, K
FUKUI, H
SHIRAKAWA, I
ISHIKAWA, E
KOBAYASHI, K
KATUNUMA, N
机构
[1] TOKUSHIMA UNIV, SCH MED, INST ENZYME RES, DEPT ENZYME CHEM, TOKUSHIMA 770, JAPAN
[2] MED COLL MIYAZAKI, DEPT BIOCHEM, KIYOTAKE, MIYAZAKI 88916, JAPAN
关键词
D O I
10.1093/oxfordjournals.jbchem.a131354
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A macromolecular antigen, ornithine .delta.-aminotransferase [EC 2.6.1.13] from rat liver (OAT) was assayed by the sandwich procedure using a rabbit (anti-OAT) Fab''-.beta.-D-galactosidase complex and rabbit (anti-OAT) IgG-coupled glass rods as a solid phase. The Fab'' fragments of the rabbit (anti-OAT) IgG were conjugated with .beta.-D-galactosidase [EC 3.2.1.23] from Escherichia coli using N,N''-o-phenylenedimaleimide. The rabbit (anti-OAT) IgG was coupled to the aminoalkylsilyl glass rods using glutaraldehyde. The rabbit (anti-OAT) IgG-coupled glass rods were incubated with OAT and then with the rabbit (anti-OAT) Fab''-.beta.-D-galactosidase complex. The amount of OAT was determined from the activity of .beta.-D-galactosidase bound to the glass rods. A minimum of 0.03 f[femto = 10-15]mol of OAT could be determined by this method. Glass rods gave greater reproducibility, and were more sensitive and simpler than use of Sepharose 4B.
引用
收藏
页码:895 / 898
页数:4
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