Production of prone‐to‐aggregate proteins

Abstract

a b s t r a c tExpression of recombinant proteins in Escherichia coli (E. coli) remains the most popular and costeffective method for producing proteins in basic research and for pharmaceutical applications. Despite accumulating experience and methodologies developed over the years, production of recombinant proteins prone to aggregate in E. coli-based systems poses a major challenge in most research applications. The challenge of manufacturing these proteins for pharmaceutical applications is even greater. This review will discuss effective methods to reduce and even prevent the formation of aggregates in the course of recombinant protein production. We will focus on important steps along the production path, which include cloning, expression, purification, concentration, and storage.

Transgenic cowpeas (Vigna unguiculata L. Walp) expressing Bacillus thuringiensis Vip3Ba protein are protected against the Maruca pod borer (Maruca vitrata)

“…The high molecular bands are possibly aggregates, a phenomenon commonly reported during the overexpression of recombinant protein in E. coli (Lebendiker and Danieli 2014). This can sometimes be overcome by the use of fusion proteins and buffers or solvents to obtain stable proteins and proper protein folding (Bondos and Bicknell 2003;Sorensen and Mortensen 2005).…”

High yields of active Thermus thermophilus proline dehydrogenase are obtained using maltose‐binding protein as a solubility tag

“…MBP is widely recognized as a carrier protein for the production of soluble recombinant proteins in E. coli [34,35]. In this report, we show that MBP can efficiently assist in the production of large amounts of soluble TtProDH in E. coli.…”

Optimization of expression and purification of recombinant S1 domain of the porcine epidemic diarrhea virus spike (PEDV- S1) protein inEscherichia coli

“…Every protein folds differently depending upon its characteristics such as size, hydrophobicity, post-translational modifications, isoelectric points, etc. [26,[42][43][44]. If the rate of protein synthesis supersedes protein folding machinery, the recombinant protein will then aggregate and result in inclusion body formation [44][45][46].…”

“…As shown in Figure 8(b), lanes 8-10, PEDV-S1 was highly enriched. Therefore, eluted fractions (19)(20)(21)(22)(23)(24)(25)(26) containing relatively pure camel were pooled. After Ni-NTA chromatography, ̴ 17 mg highly enriched Ni-NTA chromatography system is a rapid and convenient purification technique.…”

“…Keeping this in mind, this study was aimed at expressing, optimizing, and producing a large quantity of pure recombinant PEDV-S1 in Escherichia coli (E. coli). We have selected E. coli to express the recombinant PEDV-S1 because it has been proven to be a very good host for the heterologous expression of recombinant proteins, and for many purposes E. coli is the best host [24][25][26]. The recent advances in physiology and genetics of the E. coli at molecular level offer a great opportunity for the rapid and economical production of recombinant proteins [27].…”