New assay for the rapid determination of plasma holotranscobalamin II levels: preliminary evaluation in cancer patients.

We describe a new and rapid assay for the measurement of plasma B12 bound to transcobalamin II (holotranscobalamin II) using the property of adsorption of the polypeptides of apotranscobalamin II and holotranscobalamin II to the hydrophobic surface of microfine glass particles. Acid-washed microfine glass was used to separate vitamin B12 bound to the glycoproteins transcobalamin I and transcobalamin III (haptocorrin or R binder) from that bound to transcobalamin II. Sephadex gel filtration separation of 57Co-labelled vitamin B12 binders confirmed that > 90% of holotranscobalamin II can be removed from plasma holohaptocorrin by adsorption to microfine glass particles. Since only holotranscobalamin II is capable of delivering vitamin B12 to metabolizing cells, plasma holotranscobalamin II content reflects the availability of B12 to cells. Use of this test in cancer patients undergoing either chemotherapy or radiation therapy revealed evidence of early negative B12 balance that in some instances was induced by the treatment itself.