Purification of Large Quantities of Biologically Active Recombinant Human Growth Hormone

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Abstract:

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

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