Procedure for Isolation of the 20,000-Dalton Variant of Human Growth Hormone

Abstract

An isolation procedure for the 20,000-dalton variant of human growth hormone has been devised to improve the yield of the final product. The improvement involved elimination of cumbersome steps that decreased yield, and modification of chromatography on DEAE-cellu-lose to provide better separation of the variant from the major form of growth hormone.

We reported (1) an isolation procedure for the 20,000-dalton variant of human growth hormone (hGH_20K) (2) which provided quite homogeneous material but the yields were not optimum. The initial steps were cumbersome and losses resulted from them. In addition, there was the problem that in order to completely remove the major 22,000-dalton form of the hormone (hGH) during the final chromatography step (DEAE-cellulose), only the leading edge of the hGH_20k−peak could be used. The trailing part of the peak was always mixed with hGH and reworking this mixture resulted in losses. We have modified the procedure so that time-consuming steps were eliminated and the final chromatography step was improved so that now the hGH_20K can be separated from hGH by a single column.