The present report describes the application of high performance liquid chromatography (HPLC) towards the analysis of chick intestinal 1, 25-dihydroxyvitamin D3 (1, 25(OH)2D3) receptors and their interaction with anti-receptor monoclonal antibodies. Hormone-labeled vitamin D3 receptors were chromatographed under high pressure on an Altec Spherogel TSK 3000 sw column. Both radioactivity and DEAE-filtration measurements of the eluted fractions indicated a high yield (91%) of 1, 25(OH)2D3 receptor. Specific binding was both displaceable with 100-fold excess radioinert hormone and dissociable with 2 mM p-chloromercuribenzene sulfonate. In contrast, when 1, 25(OH)2D3 receptors were preincubated with anti-receptor monoclonal antibodies, the receptor's elution time was significantly accelerated to that expected for a larger macromolecule. This position corresponded to that observed for either pure or internally [35S]methionine-labeled monoclonal antibody. These results suggest that HPLC analysis and resolution of both receptor and receptor-antibody complex may be of future practical value.
|Original language||English (US)|
|Number of pages||6|
|Journal||Biochemical and Biophysical Research Communications|
|State||Published - Dec 15 1982|
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology