Fully automated protein purification.
Obtaining highly purified proteins is essential to begin investigating their functional and structural properties. The steps that are typically involved in purifying proteins can include an initial capture, intermediate purification, and a final polishing step. Completing these steps can take several days and require frequent attention to ensure success. Our goal was to design automated protocols that would allow the purification of proteins with minimal operator intervention. Separate methods have been produced and tested that automate the sample loading, column washing, sample elution and peak collection steps for ion exchange, metal affinity, hydrophobic interaction, and gel filtration chromatography. These individual methods are designed to be coupled and run sequentially in any order to achieve a flexible and fully automated protein purification protocol.
Camper
DeMarco V
DV
Department of Chemistry, University of Toledo, Toledo, OH 43606, USA.
Viola
Ronald E
RE
D001223:Aspartate-Semialdehyde Dehydrogenase / Q000302:isolation & purification*; D001331:Automation; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D002853:Chromatography, Liquid / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification*; D013296:Streptococcus pneumoniae / Q000201:enzymology; D014169:Transformation, Bacterial; D014734:Vibrio cholerae / Q000201:enzymology
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D001223:Aspartate-Semialdehyde Dehydrogenase