Investigation on Cloning and Expression of Recombinant Human Parathyroid Hormone in Lab-Scale

Document Type : Research Article


1 Faculty of Chemical Engineering, Babol Noshirvani University of Technology, Babol, I.R. IRAN

2 Medical Cellular & Molecular Research Center, Golestan University of Medical Sciences, Gorgan, I.R. IRAN


Preserving the level of calcium concentration in the blood is a dynamic process that is carried out through the interaction of several organs such as the intestine, kidney, and parathyroid glands. Human parathyroid hormone (hPTH) increases blood calcium level by increasing intestinal absorption of calcium and decreasing renal excretion. Therefore, this hormone is used for the treatment of osteoporosis and diseases of the disorders in parathyroid gland function. In this research, the plasmid pUC 57 and strain of E.coli DH5α are used for cloning and the plasmid pET 32a (+) and strain of E.coli BL21 are used for protein expression. In this study, protein production was carried out in a 1 L batch Erlenmeyer and in a 10 L fed-batch mixed bioreactor. The results of this research showed that the amounts of biomass and recombinant human parathyroid hormone were about 6.5 g/l and 1.7 g/l in shake flasks after 16 h. Also, our results showed the amounts of biomass and recombinant human parathyroid hormone were about 81 g/l and 22 g/l in the fed-batch bioreactor after 12 h, respectively. Finally, the production of recombinant human parathyroid hormone was confirmed by SDS-PAGE and western blot analyses. However, in this research, the presence of a protein band in the 37 kDa region indicates the correct expression of the parathyroid hormone. According to the successful production of human parathyroid hormone in this study, as well as the advantages of Escherichia coli as a host, it seems that this method can be a good alternative to producing this valuable drug.


Main Subjects

[1] Schmidt S.R, "Fusion Protein Technologies for Biopharmaceuticals", John Wiley & Sons Inc. (2013).
[2] Huang C.J., Lin H., Yang X., Industrial Production of Recombinant Therapeutics in Escherichia Coli and its Recent Advancement, Journal of Industrial Microbiology & Biotechnology, 3: 383-399 (2012).
[3] Bieglmayer  C., Prager  G.,  Niederle B., Kinetic Analyses of Parathyroid Hormone Clearance as Measured by Three Rapid Immunoassays During Parathyroidectomy, Clinical Chemistry , 48: 1731-1738 (2002).
[4] Francis S.,"Greenspan’s Basic & Clinical Endocinology", McGraw Hill (2011).
[5] Al-Badran A.E. , Abdul-Jabbar  R.A., Extraction and Purification of Recombinant Intact Human Parathyroid Hormone (hPTH) from Bacterial Cell, Journal of Biomedical Engineering and Informatics, 3(2): 1-10 (2017).
[6] Hamedifar  H., Salamat F., Saffarion  M., Ghiasi  M., Hosseini A., Lahiji  H., Mahboudi  F. A Novel Approach for High Level Expression of Soluble Recombinant Human Parathyroid Hormone(rhPTH 1-34) in Escherichia coli. Avicenna Journal of Medical Biotechnology , 5(3): 193-201.(2013).
 [7] Lövblad, K. O., Essig  M. Head and Neck Imaging. European Radiology Supplements, 2(1): 18-35 (2008).
[8] Vasicek, T.J., McDevitt  B.E., Freeman M.W., Fennick  B.J., Hendy  G.N., Potts  J. T.J., Kronenberg  H.M., Nucleotide Sequence of the Human Parathyriod Hormone Gene,  Proc Natl Acad Sci, 80: 2127-2131(1983).
[9] Gardella  T.J., Rubin  D., Abou- Samra  A.B., Keutmann H.T ., Potts  J.T., Kronenberg  H.M., Nussbaum S. R., Expression of Human Parathyroid Hormone-(1-84) in Escherchia Coli as a Factor X-Cleavable Fusion Protein. Journal of Biological Chemistry, 265:15854-15859 (1990).
[10] Guo  Q. R., Wei  D.-Z., Tong W. Y., Partial Purification oh Human Parathyroid Hormone 1-84 as a Thioredoxin Fusion form in Recombinant Escherichia Coli by Thermoosmotic Shock. Protein Expression and Purification, 49 (1): 32-8 (2006).
[11] Liu Q., Lin  J., Liu M., Tao X.,Wei  D., Ma X., Yang S., Large Scale Preparation of Recombinant Human Parathyriod Hormone 1-84 from Escherichia Coli, Protein Expression and Purification, 54(2): 212-219 (2007).
[13] Wang W., Tang  W.,Yan M., He K.,Yang  L., Jiang  L., Li H. A Bicistronic Expression Strategy for Large Scale Expression and Purification of Full-Length Recombinant Human Parathyroid Hormone for Osteoporosis Therapy, Protein Expression and Purification , 69(2): 178-85(2010).
[14] Bradford M.M. A Rapid and Sensitive Method for the Quantitation of Microgram of 1976 Bradford M.M Protein Utilizing the Principle of Protein-Dye Binding, Analytical Biochemistry, 72: 248-254 (1976).
[15] خوانچه زر س.،  هاشمی نجف آبادی س.،  محمدیان ج.، خلیل زاده ر،  اسفندیار س.، بهینه سازی شرایط کشت باکتری اشرشیا کولی برای اصلاح تولید قطعه نوترکیب باکتریورودوپسین، نشریه شیمی و مهندسی شیمی ایران، (2)32، 93 تا 101 (1392).