An osmolyte-based micro-volume ultrafiltration technique

Abstract

This paper discusses a novel, simple, and inexpensive micro-volume ultrafiltration technique for protein concentration, desalting, buffer exchange, and size-based protein purification. The technique is suitable for processing protein samples in a high-throughput mode. It utilizes a combination of capillary action, and osmosis for drawing water and other permeable species from a micro-volume sample droplet applied on the surface of an ultrafiltration membrane. A macromolecule coated on the permeate side of the membrane functions as the osmolyte. The action of the osmolyte could, if required, be augmented by adding a supersorbent polymer layer over the osmolyte. The mildly hydrophobic surface of the polymeric ultrafiltration membrane used in this study minimized sample droplet spreading, thus making it easy to recover the retained material after separation, without sample interference and cross-contamination. High protein recoveries were observed in the micro-volume ultrafiltration experiments described in the paper.