Preparative Purification of Recombinant Proteins: Current Status and Future Trends

Abstract

Advances in fermentation technologies have resulted in the production of increased yields of proteins of economic, biopharmaceutical, and medicinal importance. Consequently, there is an absolute requirement for the development of rapid, cost-effective methodologies which facilitate the purification of such products in the absence of contaminants, such as superfluous proteins and endotoxins. Here, we provide a comprehensive overview of a selection of key purification methodologies currently being applied in both academic and industrial settings and discuss how innovative and effective protocols such as aqueous two-phase partitioning, membrane chromatography, and high-performance tangential flow filtration may be applied independently of or in conjunction with more traditional protocols for downstream processing applications.

Biopharmaceuticals from microorganisms: from production to purification

“…This class of chromatography is probably the only technique currently available that is capable of addressing key issues in high-throughput proteomics and scale-up 62 . The most common example of an affinity process is protein-A chromatography, which has been applied for over a decade in industrial and academic settings for the capture and purification of antibodies 60 . Similarly, protein-L may possibly come to play a role in antibody fragments purification 59 .…”

Proteome analysis in the assessment of ageing

“…While the separation principle of ion-exchange and size-exclusion chromatography techniques is based respectively on charge and size properties, affinity and hydrophobic interaction chromatography on the other hand, rely on specific biochemical or hydrophobic interactions (Saraswat et al, 2013). The most popular one-dimensional chromatographic technique in proteomics is reverse-phase liquid chromatography (RP-LC) due to its high resolution and suitability with MS online coupling (Fröhlich and Arnold, 2006;Zhang et al, 2014).…”

Inteins—A Focus on the Biotechnological Applications of Splicing-Promoting Proteins

“…Epitope tags flank the protein sequence (either at the N-or the C-terminal region), facilitating purification by exploiting the affinity of the tag for a specific matrix. One of the most commonly used tags is the hexa-histidine tag [37] [39] [40]. Such affinity-based purification protocols generally require only a single step to obtain high yields of highly purified proteins [10] [39] [41] [42].…”

“…Furthermore, such purification protocols can easily be adapted for large scales, making them very convenient for industrial applications [10] [37] [39]. A drawback of this approach is that the tag generally needs to be removed at the end of the purification, a step that is frequently expensive and leads to loss of pure material [37] [39] [43]. This step is critical if the target protein is to be used as a commercial product in the food or pharmaceutical industry.…”