Optimization of a recombinant human growth hormone purification process using quality by design

Abstract

This work describes a strategy to optimize a downstream processing of a recombinant human growth hormone (rhGH) by incorporating a quality by design approach toward meeting higher quality specifications. The optimized process minimized the presence of impurities and degradation by-products during manufacturing by the establishment of in-process controls. Capillary zone electrophoresis, reverse phase, and size-exclusion chromatographies were used as analytical techniques to establish new critical process parameters for the solubilization, capture, and intermediate purification steps aiming to maintain rhGH quality by complying with pharmacopeial specifications. The results indicated that the implemented improvements in the process allowed the optimization of the specific recovery and purification of rhGH without compromising its quality. In addition, this optimization facilitated the stringent removal of the remaining impurities in further polishing stages, as demonstrated by the analysis of the obtained active pharmaceutical ingredient.

Overexpression, overproduction, purification, and characterization of rhGH in Escherichia coli

“…Divalent cation (magnesium sulfate) supplements lead to an increase in cell growth and further production of specific proteins [1, 9, 11, 30, 31]. Ethanol acts as a chemical chaperone and an inducing element for molecular chaperone synthesis by bacterial cells.…”

“…The human growth hormone (hGH) is a polypeptide chain encompassing 191 amino acids with molecular weight 22 kDa and contains two disulfide bonds, synthesized in somatotrophs in the pituitary gland. hGH is one of the most important hormones in the human body because of its pivotal role in a diversity of biological functions, including the control of cell division and metabolism [1–5]. hGH is currently used for the treatment of chronic kidney failure, short stature, Turner syndrome, Prader‐Willi, and SHOX.…”