Extraction of monoclonal antibody fragments type scFv by aqueous two-phase micellar systems

Abstract

Owing to a high percentage (70-80% correspond to the separation and purification steps) of the costs to obtain biotechnological products, it is necessary to optimize some methodologies to ensure a good cost/benefit relationship. Nowadays, the micellar aqueous two-phases systems are considered as an appropriated bio-separating technology because their properties are suitable to maintain the macromolecule's integrity and also allow regulating the experimental conditions enabling a selective extraction. In this project, the liquid-liquid extraction by micellar aqueous two-phase systems, composed by Triton X-114/ buffer McIlvaine, will be studied as a method to separate and to purify scFv antibodies from Pichia pastoris cultures. The effect of the system's components on the antibody integrity will be studied as well as the effect of different variables as pH, temperature, surfactant concentration, salts concentration and the presence of ligand molecules over the partition and purification of the scFv antibody. Finally, the best experimental conditions will be selective to apply a 1000-times scaling in order to test their potential for industrial application. (AU)