title: Isolation of ribosomes by chromatography.
abstract: Mixed-mode chromatography on cysteine-SulfoLink resin efficiently separates ribosomes from cell lysates and is particularly effective at rapidly removing endogenous proteases and nucleases, resulting in ribosomes of improved purity, integrity, and activity. Binding occurs partly by anion exchange of the RNA of the ribosomes, so that cells must be lysed in a buffer of moderate ionic strength (conductivity no more than 20 mS for chromatography of bacterial ribosomes) without any highly charged additives (e.g., heparin, which is used to inhibit RNases in yeast). A robust protocol for Escherichia coli is given here as an example.
authors.affiliation: Primary Pharmacology Group, Pfizer Global Research and Development, Groton, Connecticut 06340.
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Maguire
mesh_terms: D002845:Chromatography / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism*; D012260:Ribonucleases / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*
chemical_list: D012260:Ribonucleases