606e Selective Extraction of Recombinant Proteins by Multiple-Affinity Two-Phase Partitioning in Microchannels

Robert J. Meagher, Yooli K. Light, and Anup K. Singh. Sandia National Laboratories, P.O. Box 969, MS 9292, Livermore, CA 94551

We have demonstrated purification of proteins in a simple aqueous two-phase extraction process in a microfluidic device. The laminar flows inherent to microchannels allows us to perform a binary split of a complex cell lysate sample, in an open channel with no chromatography support and no moving parts. This mild process allows recovery of functional proteins with a modest increase in purity. Aromatic-rich fusion tags are used to drive partitioning of enzymes in a generic PEG-salt two-phase system. Addition of affinity ligands to the PEG phase allows us to exploit other popular fusion tags, such as polyhistidine tags and GST-tags.