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Protein Purification

Proteins, Peptides and Amino Acids

  1. Richard R. Burgess

Published Online: 15 SEP 2006

DOI: 10.1002/3527600906.mcb.200400065

Reviews in Cell Biology and Molecular Medicine

Reviews in Cell Biology and Molecular Medicine

How to Cite

Burgess, R. R. 2006. Protein Purification. Reviews in Cell Biology and Molecular Medicine. .

Author Information

  1. University of Wisconsin, Madison, WI, USA

Publication History

  1. Published Online: 15 SEP 2006

Abstract

Protein purification is the process of separating a given protein from all the other proteins, nucleic acids, polysaccharides, lipids, metabolites, and other small molecules in a cell extract. This purification is done ideally with reasonable speed, yield, final purity, and economy, while retaining the biological activity and chemical and structural integrity. A purification scheme involves a series of fractionation steps that exploit the many differences in the properties of the protein of interest and the other components in the mixture. Design of such fractionation steps must take into account the molecular forces that determine molecular interactions between proteins and various materials used in purification. Care must be taken to prevent inactivation, degradation or absorptive losses during the purification. In recent years, most researchers purify recombinant proteins whose genes have been cloned into an expression vector and expressed in a suitable expression host organism, often the bacterium Escherichia coli . The use of purification tags to aid in affinity purification is common. Sometimes, the overproduced protein is found as an insoluble inclusion body that must be isolated, solubilized by denaturants, and then refolded.

Keywords:

  • Fractionation;
  • Inclusion Body;
  • Protein;
  • Protein Refolding