Engineering of Therapeutic Proteins Production in Escherichia coli

Abstract

Low cost and simplicity of cultivating bacteria make the E. coli expression system a preferable choice for production of therapeutic proteins both on a lab scale and in industry. In addition straightforward recombinant DNA technology offers engineering tools to produce protein molecules with modified features. The lack of posttranslational modification mechanisms in bacterial cells such as glycosylation, proteolytic protein maturation or limited capacity for formation of disulfide bridges may, to a certain extent, be overcome with protein engineering. Protein engineering is also often employed to improve protein stability or to modulate its biological action. More sophisticated modifications may be achieved by genetic fusions of two proteins. This article presents a variety of examples of genetic engineering of therapeutic proteins. It emphasizes the importance of designing a construct without any unnecessary amino acid residues.

Top-down proteomic analysis of protein pharmaceuticals, mixtures of protein complexes, and ribosomal protein extracts by capillary zone electrophoresis-mass spectrometry

“…Recombinant DNA technology has enabled the production of human proteins in host organisms such as bacteria, resulting in structural modifications for enhanced biological activity and large scale production [8]. Recombinant DNA technology, first developed in the 1970's, initiated a revolution in the pharmaceutical industry and facilitated the development and implementation of proteins as pharmaceutical products, termed "biologics" [9]. Since the introduction of the first biologic in the US in the early 1980's, over 90 commercial biologics have been FDA approved [10].…”

DEVELOPMENT OF A QUALIFICATION PROCEDURE FOR METHIONINE FORM OF INTERFERON ALFA-2b STANDARD TO CONFIRM ITS AUTHENTICITY BY MEANS OF A PEPTIDE MAPPING METHOD

“…Поскольку синтез белка рибосомой бакте-риальной клетки инициируется АТГ-кодоном и N-конец бактериального белка представлен формилметионином, а эффективная экспрессия рекомбинантного белка в системах прокариот приводит к накоплению в клетках большого ко-личества белка, в результате чего отщепление формилметионина идет неэффективно, реком-бинантный белок может содержать дополнитель-ный метионин на N-конце молекулы [8,12].…”

Qualitative Process Understanding Tools within Bioprocessing: A Case Study

“…The model process chosen, the production of insulin from E. coli, is a complex process, which can be carried out using two methods [16]. Either the chains could be synthesised separately and mixed, reduced and reoxidised after purification [17].…”