A radioimmunoassay for phosphorylated forms of vitamin B6.
Hapten-specific antibodies are found in rabbit serum afterimmunization of the animals with phosphopyridoxyl-bovine serum albumin. These were purified approximately 80-fold by salt fractionation and affinity chromatography on phosphopyridoxyl-aminoethyl cellulose. The purified hapten-specific antibodies were covalently coupled to Immunobeads. The antibody-Immunobead preparation binds [3H]pyridoxine phosphate. This provides the basis for a radioimmunoassay for vitamin B6 derivatives. [3H]Pyridoxine phosphate is displaced from the matrix-attached antibodies by pyridoxine phosphate, pyridoxamine phosphate, and pyridoxal phosphate, but is not displaced to any significant extent by pyridoxine, pyridoxamine, pyridoxal, or 5'-deoxypyridoxal. The antibody-Immunobead preparation may be recycled for reutilization in radioimmunoassay procedures. The sensitivity of the assay is in the picomole range.
D000818:Animals; D000918:Antibody Specificity; D002417:Cattle; D002499:Centrifugation, Density Gradient; D010766:Phosphorylation; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives / Q000276:immunology; D011733:Pyridoxamine / Q000276:immunology; D011736:Pyridoxine; D011817:Rabbits; D011863:Radioimmunoassay; D012710:Serum Albumin, Bovine / Q000276:immunology
D012710:Serum Albumin, Bovine; D011732:Pyridoxal Phosphate; D011733:Pyridoxamine; D011736:Pyridoxine; C024593:pyridoxine 5-phosphate