Preparation of steroid radioligands for ultrafiltration assays by a solid-phase transport globulin method using concanavalin A-Sepharose
| Autor: | Bruce C. Nisula, Ryoji Hiramatsu |
|---|---|
| Rok vydání: | 1985 |
| Předmět: |
Male
Hydrocortisone Globulin medicine.medical_treatment Clinical Biochemistry Ultrafiltration Ligands Tritium Biochemistry Steroid Endocrinology medicine Radioligand Humans Testosterone Molecular Biology Pharmacology Chromatography biology Chemistry Sepharose Binding protein Organic Chemistry Diafiltration Steroid hormone Concanavalin A biology.protein Female Serum Globulins Steroids Protein Binding |
| Zdroj: | Steroids. 45:73-89 |
| ISSN: | 0039-128X |
| DOI: | 10.1016/0039-128x(85)90066-2 |
| Popis: | Concanavalin A-Sepharose (Con A-Sepharose) was applied to separate non-protein-bound and albumin-bound radioactive impurities from steroid radioligands. Con A-Sepharose gel, plasma, and steroid radioligand were mixed, incubated, and then washed with buffer. This method was compared with an affinity diafiltration method which separates non-protein-bound radioactive impurities with a filter membrane. /sup 3/H-Water and /sup 3/H-estrone sulfate, chosen to serve as molecules representative of non-protein-bound and albumin-bound impurities, were removed quite effectively by the Con A-Sepharose method, while 85% of /sup 3/H-estrone sulfate could not be removed by the diafiltration method. Plasma unbound cortisol (F) and testosterone (T) values determined by ultrafiltration using /sup 3/H-F and /sup 3/H-T prepared by the Con A-Sepharose method were significantly lower than those determined using the radioligands unprocessed or prepared by the diafiltration method. The whole procedure of the Con A-Sepharose method takes only 3-4 hours. This method is a simple, rapid, and effective technique for preparation of steroid radioligands for plasma protein binding studies. |
| Databáze: | OpenAIRE |
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