Purification of Large Quantities of Biologically Active Recombinant Human Growth Hormone

Abstract

Production and purification of human growth hormone using a simple method was studied in two recombinant<br />Escherichia coli, D7-5 and C27-2 strains. The r-hGH was expressed in the form of inclusion body in a batch<br />fermentation process and purified to 99% purity using a procedure based on acid precipitation of the host<br />derived proteins and other impurities. The effect of the pH and host strain on purification of the r-hGH and<br />efficiency of the procedure were evaluated. It was found that the optimum pH for precipitation of the host<br />derived proteins was 4.9. The procedure was suitable for r-hGH purification from D7-5 stain but not from the<br />other strain C27-2. The purity of > 99% and recovery of about 40% were obtained as shown by SDS-PAGE<br />and Western blot analysis. The purified r-hGH was biologically active as judged by receptor assay with<br />very low endotoxin content which could be suitable for therapeutic applications. This simple and cost effective<br />production process could be useful for large scale production of recombinant hGH from specific strains.<br /><br />