PMID: 7373061Jan 1, 1980

A radioimmunoassay for phosphorylated forms of vitamin B6

Journal of Immunological Methods
J W Thanassi, J A Cidlowski


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.


May 1, 1993·Biomedical Chromatography : BMC·S KuriokaM Matsuda

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