Model of protein isoforms analysis by aqueous two-phase systems: Methodology importance in clinical biochemistry and biopharmaceutical production

Abstract

Aqueous phase partitioning has a long history of applications to the analytical characterization of biomolecules. However, process applications have attracted the most interest in biotechnology where it has become widely recognized as a cost-effective technique. The application of aqueous two-phase systems (ATPSs) has been demonstrated in many cases including a number of industrial applications with excellent levels of purity and yield. This type of separation and purification system has also been successfully used for the separation of virus and virus-like particles. The advantage of this technique is that it may be used to monitor the aforementioned changes for purified proteins as well as for proteins in biological fluids, and that it is readily adaptable to automated high-throughput screening. However, the wide use of this technology has been diminished by the lack of a clear understanding of the factors and mechanisms that govern the behavior of proteins in these systems. It has prevented the development of analytical models that assist the rational design of these systems. This work has revised the development of ATPSs (preparation and sampling techniques), and also highlighted the knowledge gap in the ATPS.