Expression and Purification of C-Peptide Containing Insulin Using Pichia pastoris Expression System
Joint Authors
Baeshen, Nabih A.
Baeshen, Mohammed N.
Bouback, Thamer A. F.
Alzubaidi, Mubarak A.
Bora, Roop S.
Alotaibi, Mohammed A. T.
Alabbas, Omar T. O.
Alshahrani, Sultan M.
Aljohani, Ahmed A. M.
Munshi, Rayan A. A.
Al-Hejin, Ahmed
Redwan, Elrashdy M.
Ramadan, Hassan A. I.
Saini, Kulvinder S.
Ahmed, Mohamed Morsi M.
Source
Issue
Vol. 2016, Issue 2016 (31 Dec. 2016), pp.1-7, 7 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2016-08-04
Country of Publication
Egypt
No. of Pages
7
Main Subjects
Abstract EN
Increase in the incidence of Insulin Dependent Diabetes Mellitus (IDDM) among people from developed and developing countries has created a large global market for insulin.
Moreover, exploration of new methods for insulin delivery including oral or inhalation route which require very high doses would further increase the demand of cost-effective recombinant insulin.
Various bacterial and yeast strains have been optimized to overproduce important biopharmaceuticals.
One of the approaches we have taken is the production of recombinant human insulin along with C-peptide in yeast Pichia pastoris.
We procured a cDNA clone of insulin from Origene Inc., USA.
Insulin cDNA was PCR amplified and cloned into yeast vector pPICZ-α.
Cloned insulin cDNA was confirmed by restriction analysis and DNA sequencing.
pPICZ-α-insulin clone was transformed into Pichia pastoris SuperMan 5 strain.
Several Zeocin resistant clones were obtained and integration of insulin cDNA in Pichia genome was confirmed by PCR using insulin specific primers.
Expression of insulin in Pichia clones was confirmed by ELISA, SDS-PAGE, and Western blot analysis.
In vivo efficacy studies in streptozotocin induced diabetic mice confirmed the activity of recombinant insulin.
In conclusion, a biologically active human proinsulin along with C-peptide was expressed at high level using Pichia pastoris expression system.
American Psychological Association (APA)
Baeshen, Mohammed N.& Bouback, Thamer A. F.& Alzubaidi, Mubarak A.& Bora, Roop S.& Alotaibi, Mohammed A. T.& Alabbas, Omar T. O.…[et al.]. 2016. Expression and Purification of C-Peptide Containing Insulin Using Pichia pastoris Expression System. BioMed Research International،Vol. 2016, no. 2016, pp.1-7.
https://search.emarefa.net/detail/BIM-1097341
Modern Language Association (MLA)
Baeshen, Mohammed N.…[et al.]. Expression and Purification of C-Peptide Containing Insulin Using Pichia pastoris Expression System. BioMed Research International No. 2016 (2016), pp.1-7.
https://search.emarefa.net/detail/BIM-1097341
American Medical Association (AMA)
Baeshen, Mohammed N.& Bouback, Thamer A. F.& Alzubaidi, Mubarak A.& Bora, Roop S.& Alotaibi, Mohammed A. T.& Alabbas, Omar T. O.…[et al.]. Expression and Purification of C-Peptide Containing Insulin Using Pichia pastoris Expression System. BioMed Research International. 2016. Vol. 2016, no. 2016, pp.1-7.
https://search.emarefa.net/detail/BIM-1097341
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1097341