A new radioassay for serum vitamin B12 determination using chicken serum as a binder and an anion exchange resin column separation.

We present a new method for the determination of serum vitamin B12 concentration by competitive protein binding using 57Co labeled vitamin B12, chicken serum (CS) as the binding protein, and a sephadex column separation. We placed a mixture of extracted sample, labeled B12, and CS binder above an A-25 sephadex column, and after 1 hour incubation at room temperature drained the column and washed it twice with buffer. We counted the free fraction remaining in each column. Within run C.V.'s for low and high values in the normal range were 10.6% and 6.03%. The between run C.V.'s were 7.7% and 7.5% respectively. The values obtained for 95 clinical specimens correlated well with those for a reference method.