Three-phase partitioning (TPP) is rapidly developing as a novel bioseparation technique for the separation and purification of biomolecules. TPP has been applied in a wide range of applications including enzyme stability and enhancement of its catalytic activity. The partitioning into three phases is mainly dependent on the concentration of alcohol and salt used. TPP provides high enzyme recovery and can be utilized along with external techniques such as ultrasound, microwave assisted, microaffinity ligand-facilitated and also ionic-liquid based. This technique has attracted interest in the large scale recovery of proteins from crude feedstocks or fermentation broths. In this review, the basic principles, refolding of proteins using TPP, key design variables of TPP, types of TPP, applications of TPP in food industry as well as the challenge of TPP were analyzed. The work presented in this review will be beneficial for further researches in TPP or related separation techniques.

三相分配（TPP）作为一种用于分离和纯化生物分子的新型生物分离技术正在迅速发展。TPP已被广泛应用，包括酶的稳定性和增强其催化活性。分为三个阶段主要取决于所用酒精和盐的浓度。TPP可提供较高的酶回收率，并可与外部技术（例如超声，微波辅助，微亲和配体促进的以及基于离子液体的技术）一起使用。这项技术引起了人们对从粗原料或发酵液中大规模回收蛋白质的兴趣。在这篇综述中，基本原理，使用TPP进行蛋白质折叠，TPP的关键设计变量，TPP的类型，分析了TPP在食品工业中的应用以及TPP面临的挑战。这篇综述中的工作将有利于TPP或相关分离技术的进一步研究。