Preparation and Characterization of a Novel Recombinant Human Parathyroid Hormone (1 – 34) Analog (Gly1-Gln26-rhPTH(1-34)) with Enhanced Biological Activity

Title: Preparation and Characterization of a Novel Recombinant Human Parathyroid Hormone (1 – 34) Analog (Gly1-Gln26-rhPTH(1-34)) with Enhanced Biological Activity

Volume: 15 Issue: 8

Author(s): Xiao-Chao Xu, Shao-Dong Zhong, Fan Kai, Ling-Rui Li, Chao Liu, Bo Liu and Jin-Ku Bao

Affiliation:

Keywords: Gly1-Gln26-rhPTH(1-34), fusion protein, purification, cAMP, BMD, osteoporosis

Abstract: A recombinant human parathyroid hormone (rhPTH) fragment (Gly1-Gln26-rhPTH(1-34)) which contains two amino acids substitutions (Gly1 and Gln26) was acquired through Escherichia coli expression system using a soluble fusion protein strategy. The soluble fusion protein MBP-Gly1-Gln26-rhPTH(1-34) was harvested after purification by Phenyl-Sepharose F.F and Q-Sepharose F.F chromatographies. Following tobacco etch virus (TEV) protease cleavage and further purification by SP-Sepharose F.F chromatography, 30.8 mg/L Gly1-Gln26-rhPTH(1-34) without tag was obtained with high purity up to 99%. Cyclic AMP (cAMP) stimulation assay suggested that Gly1-Gln26-rhPTH(1-34) could increase the biological activity by up to 13.89% and 6.34%. After daily subcutaneous injection (for 13 weeks) of 5, 10 and 20 μg of Gly1-Gln26-rhPTH(1-34)/1000g body weight, the mean Bone Material Density (BMD) of ovariectomized (OVXed) rats increased to 7.95 – 30.54% and 1.98 – 23.32%, compared to control-vehicle group (OVX, P < 0.001) and sham- operated group (SHAM, P < 0.01), respectively.

Cite this article as:

Xu Xiao-Chao, Zhong Shao-Dong, Kai Fan, Li Ling-Rui, Liu Chao, Liu Bo and Bao Jin-Ku, Preparation and Characterization of a Novel Recombinant Human Parathyroid Hormone (1 – 34) Analog (Gly1-Gln26-rhPTH(1-34)) with Enhanced Biological Activity, Protein & Peptide Letters 2008; 15 (8) . https://dx.doi.org/10.2174/092986608785203773