title: Differential Precipitation and Solubilization of Proteins.
abstract: Differential protein precipitation is a rapid and economical step in protein purification and is based on exploiting the inherent physicochemical properties of the polypeptide. Precipitation of recombinant proteins, lysed from the host cell, is commonly used to concentrate the protein of choice before further polishing steps with more selective purification columns (e.g., His-Tag, Size Exclusion, etc.). Recombinant proteins can also precipitate naturally as inclusion bodies due to various influences during overexpression in the host cell. Although this phenomenon permits easier initial separation from native proteins, these inclusion bodies must carefully be differentially solubilized so as to reform functional, correctly folded proteins. Here, appropriate bioinformatics tools to aid in understanding a protein's propensity to aggregate and solubilize are explored as a backdrop for a typical protein extraction, precipitation, and selective resolubilization procedure, based on a recombinantly expressed protein.
authors.affiliation: School of Food Science and Environmental Health, Dublin Institute of Technology, Cathal Brugha Street, Dublin 1, Republic of Ireland. barry.ryan@dit.ie.
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Ryan
authors.affiliation: School of Food Science and Environmental Health, Dublin Institute of Technology, Cathal Brugha Street, Dublin 1, Republic of Ireland.
authors.forename: Gemma K
authors.initials: GK
authors.lastname: Kinsella
mesh_terms: D011232:Chemical Precipitation; D019295:Computational Biology / Q000379:methods; D030562:Databases, Protein; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D012995:Solubility; D064878:Web Browser
chemical_list: D011506:Proteins; D011994:Recombinant Proteins
keywords: Ammonium sulfate precipitation; Bioinformatics; Inclusion body solubilization; Protein refolding; Three phase precipitation; Trichloroacetic acid precipitation