Bioluminescent enzyme immunoassay for progesterone using monoclonal antibodies and glucose-6-phosphate dehydrogenase labels
✍ Scribed by Brochu, Michèle ;Carrié, Marie-Luce ;Térouanne, Béatrice ;Bélanger, Alain ;Nicolas, Jean-Claude
- Publisher
- John Wiley and Sons
- Year
- 1987
- Weight
- 577 KB
- Volume
- 1
- Category
- Article
- ISSN
- 0884-3996
No coin nor oath required. For personal study only.
✦ Synopsis
Bacterial luciferase, NAD( P): FMN oxidoreductase and anti-mouse immunoglobulin were co-immobilized on Sepharose 4B. This reagent together with a progesterone glucose-6phosphate dehydrogenase conjugate and various anti-progesterone monoclonal antibodies was used t o develop a non-separation bioluminescent immunoassay for progesterone. This monoclonal antibody based assay was sensitive and reliable and using the tracer progesterone-I 1-acetate-glucose-6-phosphate dehydrogenase, the majority of the monoclonal antibodies give a better sensitivity with this enzymatic tracer than that obtained with an iodinated tracer.
In a second assay design progesterone-glutathione was co-immobilized with bacterial luciferase and NAD(P): F M N oxidoreductase on Sepharose 4B and three monoclonal antibodies were labelled with glucose-6-phosphate dehydrogenase. With aqueous progesterone standards, this assay gave comparable sensitivity t o the bioluminescent enzyme immunoassay using the second antibody immunoadsorbant and t o an RIA but was unsuitable for plasma samples.