Specific purification of a single protein from a cell broth mixture using molecularly imprinted membranes for the biopharmaceutical industry

Abstract

A surface imprinting method is presented herein for the development of a highly selective yet highly permeable molecularly imprinted membrane for protein separation and purification. The resultant protein imprinted membrane was shown to exhibit great potential for the efficient separation of the template protein from a binary mixture and a cell lysate solution, while maintaining high transport flux for complementary molecules. Bovine Serum Albumin (BSA) and Lysozyme (Lys) were individually immobilized on a cellulose acetate membrane as template molecules. In situ surface crosslinking polymerization was then used for protein imprinting on the membrane for a controlled duration. Both membranes showed high adsorption capacity towards template proteins in the competitive batch rebinding tests. In addition, the adsorption capacity could be greatly enhanced in a continuous permeation procedure, where the resultant membrane specifically adsorbed the template protein for more than 40 h. Moreover, this is the first report of purification of a specific protein from the cell broth mixture using a molecularly imprinted membrane. The protein imprinted membrane enables the transport of multiple non-template proteins with high permeation rate in a complex system, thus opening the way for high efficiency protein separation at a low cost for the industry.