Enzyme immunoassay for conjugated 7alpha-hydroxy-3-oxo-4-cholenoic acid in human urine.
A microplate enzyme immunoassay (EIA) was developed for the measurement of glycine- and taurine-conjugated 7alpha-hydroxy-3-oxo-4-cholenoic acids (CDCA-delta4-3-one) in human urine. The antiserum was prepared by immunizing rabbits with N-(7alpha-hydroxy-3-oxo-4-cholen-24-oyl)-3-aminopropionic acid--bovine serum albumin conjugate. A colorimetric EIA was established using horseradish peroxidase-labeled antigen having a shorter bridge length than that of the immunogen, and 3, 3', 5, 5'-tetramethylbenzidine /hydrogen peroxide for the measurement of the enzyme activity. The reactivities of the antiserum for glycine and taurine conjugates of CDCA-delta4-3-one was almost the same. The specificity of the antiserum was investigated by determining the cross-reactivities of various bile acids and related compounds. An appropriate dose-response curve for conjugated CDCA-delta4-3-one was obtained in the range of 0.05-10 ng/well. This method was used for direct analysis of conjugated CDCA-delta4-3-one in urine of healthy infants and patients with liver diseases.
Yoshimura
T
T
Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Japan.
Taniguchi
T
T
Kobayashi
D
D
Komiya
T
T
Murai
T
T
Kimura
A
A
Kurosawa
T
T
Tohma
M
M
D002635:Chenodeoxycholic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000652:urine*; D003429:Cross Reactions; D006241:Haptens / Q000737:chemistry; D006801:Humans; D007106:Immune Sera; D007124:Immunoenzyme Techniques; D019906:Nuclear Magnetic Resonance, Biomolecular; D012680:Sensitivity and Specificity; D013055:Spectrophotometry, Infrared
D006241:Haptens; D007106:Immune Sera; D002635:Chenodeoxycholic Acid; C003019:3-oxo-7-hydroxychol-4-enoic acid