The Use of Affinity Tags to Overcome Obstacles in Recombinant Protein Expression and Purification

Title:The Use of Affinity Tags to Overcome Obstacles in Recombinant Protein Expression and Purification

Volume: 22 Issue: 10

Author(s): Chinthaka Amarasinghe and Jian-Ping Jin

Affiliation:

Keywords: Affinity chromatography, affinity tag, epitope tag, Escherichia coli, fusion protein, proteases, protein expression, purification.

Abstract: Research and industrial demands for recombinant proteins continue to increase over time for their broad applications in structural and functional studies and as therapeutic agents. These applications often require large quantities of recombinant protein at desirable purity, which highlights the importance of developing and improving production approaches that provide high level expression and readily achievable purity of recombinant protein. E. coli is the most widely used host for the expression of a diverse range of proteins at low cost. However, there are common pitfalls that can severely limit the expression of exogenous proteins, such as stability, low solubility and toxicity to the host cell. To overcome these obstacles, one strategy that has found to be promising is the use of affinity tags or carrier peptide to aid in the folding of the target protein, increase solubility, lower toxicity and increase the level of expression. In the meantime, the tags and fusion proteins can be designed to facilitate affinity purification. Since the fusion protein may not exhibit the native conformation of the target protein, various strategies have been developed to remove the tag during or after purification to avoid potential complications in structural and functional studies and to obtain native biological activities. Despite extensive research and rapid development along these lines, there are unsolved problems and imperfect applications. This focused review compares and contrasts various strategies that employ affinity tags to improve bacterial expression and to facilitate purification of recombinant proteins. The pros and cons of the approaches are discussed for more effective applications and new directions of future improvement.

Cite this article as:

Amarasinghe Chinthaka and Jin Jian-Ping, The Use of Affinity Tags to Overcome Obstacles in Recombinant Protein Expression and Purification, Protein & Peptide Letters 2015; 22 (10) . https://dx.doi.org/10.2174/0929866522666150728115307