Aqueous Two-Phase System Strategies for the Recovery of Proteins from Plants

Abstract

The increasing interest of the biopharmaceutical industry to exploit plants as a commercially viable production system is demanding the development of new strategies to maximize product recovery. Aqueous two-phase systems (ATPSs) are a primary recovery technique that has shown great potential for the efficient extraction and purification of biological products. The evaluation of different system parameters upon the partitioning behavior can provide the conditions that favor the concentration of contaminants and the desired target protein in opposite phases. The protocols described provide the basic strategy to explore the use of ATPS for the isolation and partial purification of native and recombinant proteins expressed in plants.

Development of Enzyme-Based Cosmeceuticals: Studies on the Proteolytic Activity of Arthrospira platensis and Its Efficient Incorporation in a Hydrogel Formulation

“…Separation of the proteolytic activity of A. platensis lysate was achieved using an aqueous two-phase partition system (ATPS) composed by polyethylene glycol (PEG)/phosphate buffer. The optimal molecular weight of PEG, the pH and the salt concentration in the system [69] were evaluated in order to optimize the separation and purification efficiency of the method. Initially, the effect of PEG's molecular weight on separation was determined with the following conditions: (a) 27.4% (w/w) PEG 1500 (stock solution 80% (w/w)), 32.7% (w/w) phosphate pH 7.0, 30% (w/w) ddH 2 0, 10% (w/w) sample, and (b) 33.8% (w/w) PEG 3000 (stock solution 50% (w/w)), 36.2% (w/w) phosphate buffer pH 7.0, 20% (w/w) ddH 2 O, 10% (w/w) extract.…”

“…The parameters that affect the purifying efficiency of the ATPS toward the desired proteins were adjusted and evaluated in order to establish the optimal system conditions. The evaluated parameters included the molecular weight (1500 or 3300 Da) of PEG, the pH (6.5-8.5) of the aqueous phase (phosphate buffer) and the salt concentration (NaCl, 0-10% w/w) in the system [69]. The results are listed in Table 1.…”