Current affinity approaches for purification of recombinant proteins

Sahar Mahmoodi,Hasan Majdi,Nosratollah Zarghami,Mohammad Pourhassan-Moghaddam,David W Wood,Rajni Hatti Kaul

doi:10.1080/23312025.2019.1665406

Abstract

Recombinant proteins have wide applications in the development of pharmaceutical compounds, industrial applications of enzymes, and basic proteomics research. In this way, efficient production of recombinant proteins with high purity needs efficient purification methods. Various strategies have been devised to improve these proteins purification such as affinity purification and physicochemical purification methods which the affinity purification has some advantages over the others. Affinity strategies especially fusion strategies have been devised as indispensable tools for the massive parallel production, identification and purification of recombinant proteins from the host systems. These strategies facilitate commercial and industrial formulations of recombinant proteins, improve the study of protein interactions at the molecular level, and develop highly sensitive and specific bioassays. Recently, various surface-modified nanoparticles have been widely developed to enhance recovery and purification of recombinant proteins such as Hydrophobic polymer nanoparticles and Oleosin nanoparticles. In this review, we aim to discuss affinity purification technologies and address the principles, advantages, limitations and potential applications of them.

Highlights

The heterologous expression of proteins is a necessary step for the study of biological functions of genes, development of pharmaceutical compounds, industrial applications of enzymes, and basic proteomics research
The results showed a very efficient purification of all three proteins using cobalt (II) ions immobilized on carboxymethyl aspartate cross-linked agarose under near-physiological conditions
Numerous subsequent chromatographic phases are performed to obtain a relatively pure protein. These methods are timeconsuming approaches with a low yield of recovery, whereas affinity chromatography leads to high yields, reduction in the number of steps, costs and impurities (Arnau et al, 2006; Jain, 2006; Korf et al, 2005)

Summary

Introduction

The heterologous expression of proteins is a necessary step for the study of biological functions of genes, development of pharmaceutical compounds, industrial applications of enzymes, and basic proteomics research. The main advantage of using recombinant proteins is that a wealth of information is often already available about the product and major impurities, thereby simplifying detection assays, sample preparation methods, and purification strategies This information allows the rapid development of strategies for both industrial and research-scale production of specific recombinant protein targets. The economical production of recombinant proteins requires implementation of efficient purification methods as well as high-yielding expression hosts (Healthcare, 2007). To address this need, various strategies have been devised to improve the recovery rates of the desired recombinant proteins, including physicochemical purification methods and affinity purification methods (Kimple, Brill, & Pasker, 2013; Kosobokova, Skrypnik, & Kosorukov, 2016; Wingfield, 2015)

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Conclusion