Cellular engineering for therapeutic protein production: product quality, host modification, and process improvement

Abstract

Recombinant proteins offer many therapeutic advantages unavailable in traditional small molecule drugs, but the need for cellular versus chemical synthesis complicates production. Avenues for producing therapeutic biologics are continuously expanding, and developments in biochemistry, cell biology, and bioengineering fuel new discoveries that promise safer, more efficient, and cheaper drugs for consumers. Numerous approaches to express recombinant proteins exist, but Escherichia coli, Saccharomyces cerevisiae, and mammalian systems (e.g. Chinese hamster ovary cells, CHO) are the most widely utilized. Improvements to production in these hosts have focused on novel expression cassettes, cell line modifications, engineering secretion pathways, and media design. Here, we describe recent developments for improving protein production in E. coli, S. cerevisiae, and CHO systems and compare recent advancements to previous knowledge in the field. With the expanding importance and prevalence of protein therapeutics, these improvements will serve as the framework for future discoveries.

Impact of CHO Metabolism on Cell Growth and Protein Production: An Overview of Toxic and Inhibiting Metabolites and Nutrients

“…The main advantages of using CHO cells compared to other microbial or mammalian cells include the ability of these cells to perform post‐translational modifications similar to those found in human proteins, such as glycosylation, which is considered to be a critical quality attribute. The presence of an aberrant glycan profile will decrease the efficacy, affects the protein drug pharmacokinetics, and alters biological properties .…”

Applying Unconventional Secretion in Ustilago maydis for the Export of Functional Nanobodies

“…Various well-established expression systems exist, often providing high space-time yields [4,5]. These include for example bacterial systems like Escherichia coli or Bacillus subtilis in which mostly unmodified proteins are produced [6,7,8].…”

“…The yeasts Saccharomyces cerevisiae and Pichia pastoris , or filamentous fungi like Aspergilli are established workhorses mainly for generating eukaryotic proteins [9,10,11]. For production of expensive therapeutic proteins, human or CHO cell cultures are preferentially used, especially if post-translational modifications are required for protein function [5,12]. However, these cultures are more costly, slow growing, and it is elaborate and time-consuming to develop expression lines [4].…”

Editorial: Methods and advances for systems and synthetic biology, nanobiotech and medicine

“…Professor Anne Robinson and Evan Wells describe recent developments for improving protein production in Escherichia coli, Saccharomyces cerevisiae, and CHO systems and introduce the most recent advancements in the field which will serve as the framework for future discoveries [10]. The development of new cancer therapeutics is asking for efficient screening methods in cell cultures.…”