Genetic Engineering & Biotechnology News

MAY15 2017

Genetic Engineering & Biotechnology News (GEN) is the world's most widely read biotech publication. It provides the R&D community with critical information on the tools, technologies, and trends that drive the biotech industry.

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22 | MAY 15, 2017 | | Genetic Engineering & Biotechnology News Hagen Richter, Ph.D., and Ilona Koebsch, Ph.D. The success of biopharmaceuticals started about 30 years ago with the first production of recombinant human insulin (Humulin ® ) in Escherichia coli, followed by the first produc- tion of human tissue plasminogen activator (tPA) in mammalian host cells some years later. Due to the lack of glycosylation in E. coli, which is required for the biological activity of most monoclonal antibodies, the use of Chinese hamster ovary (CHO) cell lines soon became the industrial gold standard for the production of biopharmaceuticals. CHO cells possess the machinery for post-translational modifications and, in con- trast to conventional E. coli systems, make it possible to purify correctly folded and secreted proteins directly from the culture broth. Consequently, the booming demand for antibodies led to the success of CHO cells in biomanufacturing. Nevertheless, CHO-based systems still suffer from slow cell growth and thus low productivity. Moreover, process develop- ment using mammalian cells is time-con- suming, due to tedious clone screening and selection, which can take up to five months. The demand for fast, safe, and cost-effi- cient manufacturing solutions is triggered by increasing pressure on clinical development timelines and public healthcare systems. Per- sonalized medicine and biosimilars are just two examples underlining the need for in- novative expression platforms that combine high productivity, protein secretion, and rap- id process development. Esetec E. coli is a well-studied and quick-repli- cating host with a genetic system that is eas- ily manipulated. The fast-growing nature of E. coli accelerates process development, with less time spent on clonal screening, cell line development, cultivation, and testing. Common disadvantages are the laborious purification from the periplasm and refold- ing from inclusion bodies. Such constraints, however, have been conquered by the pro- prietary E. coli expression system Esetec ® (E. coli secretion technology) developed by Wacker Biotech. Safe E. coli K12 strains have been engi- neered to secrete correctly folded proteins directly into the culture medium. The unique ability to export the proteins enables purifi- cation of the product without cell disruption and results in higher yields and quality. The secretion of the target protein reduces pro- cess-related impurities, like host cell DNA and endotoxins, which need to be removed by more extensive purification in conven- tional E. coli procedures. Recent improve- ments of the Esetec technology have allowed high-level expression and secretion of pro- teins that are difficult to express. The broad range of secreted products with molecular weights of 5 to 150 kDa renders Esetec a versatile and cost-efficient alternative for any nonglycosylated biopharmaceutical. Time Is Critical Biopharmaceutical drug development re- quires several rounds of clinical testing with an increasing amount of drug substance needed. Regrettably, the failure rate of early clinical candidates is more than 90%, fuel- ing the demand for rapid and reliable pro- duction systems to cope with the increasing number of clinical studies. The long history of CHO cells for the standardized production of antibodies has helped to improve development timelines. Ideally, generic purification approaches counterbalance time-consuming cell-line de- velopment, therefore reducing the advertised time from gene to GMP-grade antibody ma- terial to 11 months. Process development and GMP manufac- turing of nonantibody products, however, re- quire at least 16 months, mostly due to higher efforts and longer process development time- lines (Figure 1). Esetec takes advantage of a fast-growing host strain and protein secre- tion, which speeds up process development and production. In total, only 12 months are required from gene to the first GMP batch, even for nonplatform products (Figure 1). By leveraging the Esetec advantage in GMP manufacturing, the typical time-in-facil- ity of a batch is just one third of that of mam- malian cell cultures. Due to shorter fermenta- tion times and based on a simulated process, batch production with Esetec is completed after seven days, while CHO cells require ap- proximately 20 days. Comprehensive Cost Analysis A comprehensive cost analysis for different expression systems is difficult, as each process varies, depending on the protein of interest. To System Was Developed to Create a Cost-Efficient Alternative to Mammalian Cell Culture Microbial Secretion via Esetec Technology Bioprocessing Tutorial Figure 1. From gene to first GMP batch—timeline comparison Esetec versus CHO. Duration of cell-line/strain selection, process development, scale-up, and GMP manufacturing is shown. Data for CHO cells is based on data of two relevant market players. The typical development and production timeline of Esetec saves up to four months compared to mammalian cell culture. Advantages of Esetec are faster strain development and the lack of development of viral depletion steps (not necessary for microbial systems). BioResearch NIH Authentication Requirement * Proceed with Confidence CellCheck ™ cell line authentication is the gold-standard solution for addressing NIH grant authentication requirements. CellCheck provides one easy step for one low price, detecting both contamination and misidentification. Order CellCheck and receive your free Authentication Guide that offers tools, tips and resources to improve your NIH grant application. To order CellCheck and your free Authentication Guide: Robyn Myles • 573 499 5722 (direct) • 800 669 0825 • *NIH notification #NOT-OD-16-011, Implementing Rigor & Transparency in NIH. © 2017 IDEXX Laboratories, Inc. All rights reserved.

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