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

BioMed Research International

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

Medicine

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