Development of a solid-phase enzyme immunoassay for ursodeoxycholic acid: application to plasma disappearance of injected ursodeoxycholic acid in the rabbit.
A bile acid disappearance test using an enzyme immunoassay for ursodeoxycholic acid (UDCA) is presented. The immunoassay employs an antiserum produced in rabbits with UDCA coupled by amide linkage to egg albumin. An antigen (UDCA)-enzyme (beta-D-galactosidase) complex was prepared by adding the N-hy...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
1979-11-01
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| Series: | Journal of Lipid Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520399971 |
| _version_ | 1823927944296267776 |
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| author | Y Maeda T Setoguchi T Katsuki E Ishikawa |
| author_facet | Y Maeda T Setoguchi T Katsuki E Ishikawa |
| author_sort | Y Maeda |
| collection | DOAJ |
| description | A bile acid disappearance test using an enzyme immunoassay for ursodeoxycholic acid (UDCA) is presented. The immunoassay employs an antiserum produced in rabbits with UDCA coupled by amide linkage to egg albumin. An antigen (UDCA)-enzyme (beta-D-galactosidase) complex was prepared by adding the N-hydroxy-succinimide ester of UDCA to beta-D-galactosidase in a molar ratio of 5000:1. The anti-UDCA serum was coupled to glass beads and a competitive reaction between bile acids and UDCA coupled to the enzyme on the glass beads was measured by determining enzyme activity. One bead was used for each test tube. Thus it was convenient to wash and transfer the bead to a fresh test tube after incubation. The procedure requires 2.5 hr at 30 degrees C for the competitive reaction and enzyme assay. Using a 1:100 dilution of anti-serum, the intensity of fluorescence of 4-methylumbelliferone produced from 4-methylumbelliferyl-beta-D-galactoside by the enzyme decreased linearly with a logarithmic increase of UDCA concentration over a range of from 0.1 to 10 pmnd taurine conjugates, and good recovery data were obtained. The development of the enzyme immunoassay using glass beads shortens analysis time; furthermore, the method makes it possible to detect obstructive jaundice in rabbits before the serum bilirubin level is elevated. |
| first_indexed | 2024-12-16T20:46:51Z |
| format | Article |
| id | doaj.art-b24606d2a98a4404b1b3019e9e17c1ea |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-16T20:46:51Z |
| publishDate | 1979-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-b24606d2a98a4404b1b3019e9e17c1ea2022-12-21T22:16:54ZengElsevierJournal of Lipid Research0022-22751979-11-01208960965Development of a solid-phase enzyme immunoassay for ursodeoxycholic acid: application to plasma disappearance of injected ursodeoxycholic acid in the rabbit.Y MaedaT SetoguchiT KatsukiE IshikawaA bile acid disappearance test using an enzyme immunoassay for ursodeoxycholic acid (UDCA) is presented. The immunoassay employs an antiserum produced in rabbits with UDCA coupled by amide linkage to egg albumin. An antigen (UDCA)-enzyme (beta-D-galactosidase) complex was prepared by adding the N-hydroxy-succinimide ester of UDCA to beta-D-galactosidase in a molar ratio of 5000:1. The anti-UDCA serum was coupled to glass beads and a competitive reaction between bile acids and UDCA coupled to the enzyme on the glass beads was measured by determining enzyme activity. One bead was used for each test tube. Thus it was convenient to wash and transfer the bead to a fresh test tube after incubation. The procedure requires 2.5 hr at 30 degrees C for the competitive reaction and enzyme assay. Using a 1:100 dilution of anti-serum, the intensity of fluorescence of 4-methylumbelliferone produced from 4-methylumbelliferyl-beta-D-galactoside by the enzyme decreased linearly with a logarithmic increase of UDCA concentration over a range of from 0.1 to 10 pmnd taurine conjugates, and good recovery data were obtained. The development of the enzyme immunoassay using glass beads shortens analysis time; furthermore, the method makes it possible to detect obstructive jaundice in rabbits before the serum bilirubin level is elevated.http://www.sciencedirect.com/science/article/pii/S0022227520399971 |
| spellingShingle | Y Maeda T Setoguchi T Katsuki E Ishikawa Development of a solid-phase enzyme immunoassay for ursodeoxycholic acid: application to plasma disappearance of injected ursodeoxycholic acid in the rabbit. Journal of Lipid Research |
| title | Development of a solid-phase enzyme immunoassay for ursodeoxycholic acid: application to plasma disappearance of injected ursodeoxycholic acid in the rabbit. |
| title_full | Development of a solid-phase enzyme immunoassay for ursodeoxycholic acid: application to plasma disappearance of injected ursodeoxycholic acid in the rabbit. |
| title_fullStr | Development of a solid-phase enzyme immunoassay for ursodeoxycholic acid: application to plasma disappearance of injected ursodeoxycholic acid in the rabbit. |
| title_full_unstemmed | Development of a solid-phase enzyme immunoassay for ursodeoxycholic acid: application to plasma disappearance of injected ursodeoxycholic acid in the rabbit. |
| title_short | Development of a solid-phase enzyme immunoassay for ursodeoxycholic acid: application to plasma disappearance of injected ursodeoxycholic acid in the rabbit. |
| title_sort | development of a solid phase enzyme immunoassay for ursodeoxycholic acid application to plasma disappearance of injected ursodeoxycholic acid in the rabbit |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520399971 |
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