A New Principle for Measurement of Cobalamin and Corrinoids, Used for Studies of Cobalamin Analogs on Serum Haptocorrin

Transcobalamin (TC) and haptocorrin (HC) are serum corrinoid-binding proteins. We developed new methods for measurement of the corrinoids bound to HC and TC. TC (n = 10) or HC (n = 138) was immunoprecipitated, and corrinoids were released by enzymatic degradation [subtilisin Carlsberg (EC 3.4.21.62)] of the binding proteins. Binding of the released corrinoids to added unsaturated TC (apoTC) or HC (apoHC) created holoTC (as measure of cobalamins) and holoHC (as measure of corrinoids). holoTC and holoHC were measured by use of ELISA. The amounts of analogs were calculated as the difference between corrinoids and cobalamins. Corrinoids extracted from HC were separated with HPLC after addition of potassium cyanide (n = 3). The corrinoid- and cobalamin-specific assays had a positive linear relation between analyte concentration and assay signal, detection limits of 8 and 4 pmol/L, and imprecision values (CV) of