Genetic engineering to enhance the selectivity of protein separations |
| |
Authors: | Charles E Glatz Clark F Ford |
| |
Institution: | (1) Department of Chemical Engineering, Iowa State University, 50011 Ames, IA;(2) Department of Food Science and Human Nutrition, Iowa State University, 50011 Ames, IA |
| |
Abstract: | The ability to recover and purify natural and recombinant proteins, and the costs of doing so remain a major task in introducing
the potential products of biotechnology. The 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. In most cases, a combination of methods
is used in sequence, and improvements in the selectivity at an early stage can enhance the effectiveness of subsequent (and
usually more costly) steps. Genetic engineering provides a means of improving the selectivity within the context of existing
separation methods.
By this strategy, improvements in selectivity are sought by bestowing a distinctive property on the protein of interest. The
primary sequence of amino acids is altered, such that the protein can be selectively removed from other components of the
multicomponent mixture in which such products are commonly found. In this article, the range of these “distinctive properties”
and their pairing with various separation methods will be reviewed. Specific examples from our work, in which a distinctive
charge is provided via a polypeptide “purification” fusion tail, will be discussed. Separation methods we have used with these
fusion proteins are precipitation, two-phase aqueous extraction, reversed micellar extraction, and ion exchange using both
resins and membranes. |
| |
Keywords: | Protein separation purification fusion precipitation ion exchange genetic engineering |
本文献已被 SpringerLink 等数据库收录! |
|