Opportunities and challenges of the tag-assisted protein purification techniques: Applications in the pharmaceutical industry

• Published in: Biotechnology advances 2020-12, Vol.45, p.107653-107653, Article 107653
• Main Authors: Mahmoudi Gomari, Mohammad, Saraygord-Afshari, Neda, Farsimadan, Marziye, Rostami, Neda, Aghamiri, Shahin, Farajollahi, Mohammad M.
• Format: Article
• Language: English
• Subjects: Aptamer; Biomimetic; Chromatography, Affinity; Drug Industry; Epitope; General-ligand bioaffinity; Nanotechnology; Peptides; Protease; Proteins; Recombinant Fusion Proteins; Recombinant proteins; Self-cleavable tags; Specific-ligand bioaffinity; Tandem affinity purification
• ISSN: 0734-9750; 1873-1899
• DOI: 10.1016/j.biotechadv.2020.107653

Tag-assisted protein purification is a method of choice for both academic researches and large-scale industrial demands. Application of the purification tags in the protein production process can help to save time and cost, but the design and application of tagged fusion proteins are challenging. An appropriate tagging strategy must provide sufficient expression yield and high purity for the final protein products while preserving their native structure and function. Thanks to the recent advances in the bioinformatics and emergence of high-throughput techniques (e.g. SEREX), many new tags are introduced to the market. A variety of interfering and non-interfering tags have currently broadened their application scope beyond the traditional use as a simple purification tool. They can take part in many biochemical and analytical features and act as solubility and protein expression enhancers, probe tracker for online visualization, detectors of post-translational modifications, and carrier-driven tags. Given the variability and growing number of the purification tags, here we reviewed the protein- and peptide-structured purification tags used in the affinity, ion-exchange, reverse phase, and immobilized metal ion affinity chromatographies. We highlighted the demand for purification tags in the pharmaceutical industry and discussed the impact of self-cleavable tags, aggregating tags, and nanotechnology on both the column-based and column-free purification techniques. •The application of tags for protein purification will reduce the cost of preparing the chromatographic columns.•A suitable tagging system may provide a sufficient expression yield and high purity of the protein product.•The introduction of the self-cleavable tags and aggregating tags have been influential in the protein purification industry.•The application of the aggregating tags in a column-free manner can facilitate higher protein expression yield.•Emerging technologies, including bioinformatics, SELEX, and nanotechnology, have helped to improve technical limitations.