Genetic Engineering for Product Using Non-Chromatographic Methods

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The ability to recover and purify natural and recombinant proteins, and the costs of doing so, remain a major task in introducing potential products for biotechnology. The available bases for separation range from specific binding onto tailored reagents to solubility and partitioning behavior governed by a mixed bag of size, charge and hydrophobicity. The separation methods considered here are precipitation, and ion-exchange using both resins and membranes. Improvements in selectivity are sought by bestowing a distinctive charge on the protein of interest such that a charge-based separation method provides selective recovery. The charge is provided using a polypeptide "purification" fusion tail introduced by placing a synthetic oligonucleotide at the end of the gene for the protein. The series of fusion proteins provides a means of studying the charge-dependent phenomena in separation methods which may be dependent on several characteristics of the protein.