Volume 10, number 1
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Effects of Different Buffers and pH on the Stability of Recombinant Human Growth Hormone

Hamid Mohammadpanah1, Hossein Rastegar2, Mohammad Reza Ramazani3 and Mahmoud Reza Jaafari1*

1Biotechnology and Nanotechnology Research Center, School of Pharmacy, Mashhad University of Medical Sciences (MUMS), Mashhad, Iran.

2Food and Drug Control Laboratories and Research Center, Ministry of Health and Medical Education, Tehran, Iran.

3Daroosazi Samen Pharmaceutical Co. Biotechnology Department, Mashhad, Iran.

DOI : http://dx.doi.org/10.13005/bbra/1110

ABSTRACT: Recombinant Human Growth hormone (rHGH) is a pharmaceutical single-chain polypeptide that contains 191 amino acids, with two disulfide bonds. The objective of this study was to determine the effects of different buffers and pH on the stability of rHGH and select the best ones for increasing the stability of rHGH in liquid solution. rHGH were dissolved at 3.33 mg/ml in phosphate (5mM and 10 mM), L-histidin (10 mM), sodium citrate (10 mM) and ammonium hydrogen carbonate (10 mM) buffers with the pH of 6.0, 6.25, 6.5 and 7.0 containing 2.5 mg/ml phenol. Stability of rHGH in the presence of different buffers and pH was evaluated by a size-exclusion chromatography (SEC) to determine the changes of the rHGH monomer to dimer, oligomer, and high molecular mass (Dimer & HM) and RP-HPLC to determine chemical decomposition and deamidation of rHGH. The effect of different pH on aggregation and precipitation, due to the agitation of rHGH solution, was evaluated by UV spectroscopy and Light Scattering Particle size analysis methods. Analysis of stability studies at 4, 25 and 37°C revealed that pH variation had a significant effect on rHGH stability. As the pH shift from 6 to 7 changes of protein to Dimer and H.M. decreased at the contrary changes of rHGH to related protein is increased. Altogether the results indicates that histidine or citrate buffer at 10 mM concentration and pH 6.25 to 6.5 provides better stability both regards to its physical and chemical stability.

KEYWORDS: Recombinant Human Growth Hormone; rHGH; Stability; Somatropin; Buffers; pH

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Mohammadpanah H, Rastegar H, Ramazani M. R, Jaafari M. R,. Effects of Different Buffers and pH on the Stability of Recombinant Human Growth Hormone. Biosci Biotechnol Res Asia 2013;10(1)
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Mohammadpanah H, Rastegar H, Ramazani M. R, Jaafari M. R,. Effects of Different Buffers and pH on the Stability of Recombinant Human Growth Hormone. Biosci Biotechnol Res Asia 2013;10(1). Available from:https://www.biotech-asia.org/?p=10302

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