Challenges and opportunities in the purification of recombinant tagged proteins

• Published in: Biotechnology advances Vol. 32; no. 2; pp. 366 - 381
• Main Authors: Pina, Ana Sofia, Lowe, Christopher R., Roque, Ana Cecília A.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier Inc 01.03.2014; Elsevier; Elsevier Inc. Published by Elsevier Inc
• Subjects: Adsorbents; Affinity; affinity chromatography; Affinity ligands; Affinity tags; Animals; Biological and medical sciences; Biotechnology; Chromatography, Affinity; Fundamental and applied biological sciences. Psychology; Fusion proteins; Humans; Ligands; Mice; Proteins; Purification; Recombinant; Recombinant Fusion Proteins - isolation & purification; Recombinant proteins; Research Review Paper; Strategy; Tags; Affinity tags; Fusion proteins; Recombinant proteins; Affinity ligands; Purification; Ligand; Affinity; Recombinant protein; Fusion protein
• ISSN: 0734-9750; 1873-1899; 1873-1899
• DOI: 10.1016/j.biotechadv.2013.12.001

The purification of recombinant proteins by affinity chromatography is one of the most efficient strategies due to the high recovery yields and purity achieved. However, this is dependent on the availability of specific affinity adsorbents for each particular target protein. The diversity of proteins to be purified augments the complexity and number of specific affinity adsorbents needed, and therefore generic platforms for the purification of recombinant proteins are appealing strategies. This justifies why genetically encoded affinity tags became so popular for recombinant protein purification, as these systems only require specific ligands for the capture of the fusion protein through a pre-defined affinity tag tail. There is a wide range of available affinity pairs “tag-ligand” combining biological or structural affinity ligands with the respective binding tags. This review gives a general overview of the well-established “tag-ligand” systems available for fusion protein purification and also explores current unconventional strategies under development. •Current and future trends for the purification of recombinant proteins•Comparison of affinity ligands for fusion protein purification•Versatile and unconventional purification strategies for fusion proteins