Isoelectric Separation of Proteins using Charged Ultrafilter Membranes with Different Functionality under Coupled Driving Forces

Abstract

A simple membrane process for the protein fractionation under coupled driving forces (pressure and potential difference) has been developed using acidic functionalized (sulphonated, carboxyleted, and phosphorylated) ultrafilter membranes, based on the interpolymer of poly(vinyl chloride) (PVC) and styrene-divinyl benzene (DVB) copolymer. Introduction of the different functional groups was confirmed by Fourier transform infrared (FTIR), CHNS analysis, and ion-exchange capacity measurements. Molecular weight cut off (MWCO) determination of these membranes suggested their ultrafilter nature, while their contact angle values showed hydrophilic characteristics. The apparent pore radius of these membranes was estimated by water permeation studies, while electro-osmotic permeation data was used for the determination of zeta potential under the operating environment. Systematic studies on the effects of pH, or nature of the charge on the casein (CAS) and lysozyme (LYS), on their adsorption characteristic using these charged ultrafilter membranes were carried out. Protein transmission (selectivity) and membrane throughput across both membranes were studied using binary mixture of protein under different gradients at pH points: 2.0, 5.0, 10.7, and 13.0. It was concluded that separation from the binary mixture of CAS−LYS of LYS at pH 5.0 (pI of CAS) using charged ultrafilter membranes was possible with high selectivity and throughput. It was observed that transmission of protein can be governed by varying the nature and extent of charge on the protein (pH) and membrane matrix, polarity of applied potential gradient with an ultrafilter membrane of given pore dimensions. In these novel processes, charge on the protein, nature and extent of the charge on the membrane interfaces, and polarity of the potential gradient all are governing the transport of a given protein across the membrane, which resulted high selectivity and membrane throughput under coupled driving forces.