Iodogen-catalysed iodination of human calcitonin and Tyr(0)-katacalcin and purification of their mono- and di-iodinated derivatives by chromatofocussing.

Synthetic calcitonin (CT) and Tyr(0)-katacalcin (tKT) were radioiodinated with Iodogen to high specific activity and with high yields. The products of the iodination procedure were chromatographed on Sephadex G-25 to remove unreacted iodide and then separated by chromatofocussing on PBE 94 (pH 9.6-6.0 for CT and pH 7.4-4.0 for tKT). Clear separation between uniodinated peptides and their mono- and di-iodinated derivatives was achieved with specific activities of 1900 and 3800 Ci/mmol for the respective mono- and di-iodinated peptides. Yields were up to 36 and 24% of mono- and di-iodinated CT and 41 and 29% for mono- and di-iodinated tKT. Our results show that Iodogen provides an effective and gentle way to iodinate peptides with high efficiency. Chromatofocussing is a simple, inexpensive and instrumentally undemanding method that can be performed without specialized chromatographic equipment and that should be applicable to a variety of different tracer preparations.