Cell factories for insulin production.

Nabih A Baeshen,Mohammed N Baeshen,Hassan A I Ramadan,Roop S Bora,Abdullah Sheikh,Mohamed Morsi M Ahmed,Elrashdy M Redwan,Kulvinder Singh Saini

doi:10.1186/s12934-014-0141-0

Abstract

The rapid increase in the number of diabetic patients globally and exploration of alternate insulin delivery methods such as inhalation or oral route that rely on higher doses, is bound to escalate the demand for recombinant insulin in near future. Current manufacturing technologies would be unable to meet the growing demand of affordable insulin due to limitation in production capacity and high production cost. Manufacturing of therapeutic recombinant proteins require an appropriate host organism with efficient machinery for posttranslational modifications and protein refolding. Recombinant human insulin has been produced predominantly using E. coli and Saccharomyces cerevisiae for therapeutic use in human. We would focus in this review, on various approaches that can be exploited to increase the production of a biologically active insulin and its analogues in E. coli and yeast. Transgenic plants are also very attractive expression system, which can be exploited to produce insulin in large quantities for therapeutic use in human. Plant-based expression system hold tremendous potential for high-capacity production of insulin in very cost-effective manner. Very high level of expression of biologically active proinsulin in seeds or leaves with long-term stability, offers a low-cost technology for both injectable as well as oral delivery of proinsulin.

Highlights

The pioneering work of Stanley Cohen and Herbert Boyer, who invented the technique of DNA cloning, signaled the birth of genetic engineering, which allowed genes to transfer among different biological species with ease [1]
Another approach involves the expression of a single chemically synthesized cDNA encoding for human proinsulin in E. coli followed by purification and subsequent excision of C-peptide by proteolytic digestion
This study clearly demonstrated that expression of insulin as oleosin fusion protein in plant allow accumulation of large amount of recombinant insulin within the seed and provide simple downstream purification by centrifugation i.e. oilbody purification

Summary

Introduction

The pioneering work of Stanley Cohen and Herbert Boyer, who invented the technique of DNA cloning, signaled the birth of genetic engineering, which allowed genes to transfer among different biological species with ease [1]. The first commercial recombinant insulin was developed for therapeutic use in human by this two-chain combination procedure [60] Another approach involves the expression of a single chemically synthesized cDNA encoding for human proinsulin in E. coli followed by purification and subsequent excision of C-peptide by proteolytic digestion. This approach was more efficient and convenient for large scale production of therapeutic insulin as compared to the two chain combination approach and has been used commercially since 1986 [60]. Insulin Detemir is another recombinant long-acting insulin analogue that was commercially produced in S. cerevisiae, developed by Novo Nordisk and approved for therapeutic use in human in 2004 by European regulatory authorities. Cairo 12311, Egypt. 4Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City for Scientific Research and Applied Technology, New Borg AL-Arab, Alexandria, Egypt

Conclusion

34. Gerngross TU

38. Ahmad B

50. Kane JF

Findings

58. De Marco A