CHEM 271 Lecture Notes - Lecture 6: Enzyme, Protein Purification, Molecular Mass
10 Jan 2018
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Document Summary
Starting with a default mixture, the requirement was to isolate protein #17. The pre-purification information given was that the maximal temperature to proceed with was of 50 c and to stay within a range of ph 4 to 10. 5 to keep the enzymatic activity stable. The purification process was done by first passing the proteins in a sephadex g-50a gel-filtration media to remove all proteins under 30 000 molecular mass. Secondly, the protein mixture left was heat treated for 120 minutes at 50 c to denature all proteins which cannot withstand that temperature. Lastly, the mixture was passed through a phenyl-sepharose 2m to 0m hydrophobic interaction chromatography to remove the hydrophobic proteins. The plan was cost-effective in that the hours were limited to the lowest amount possible while effectively purifying the protein in a minimal number of steps; there was a total of 3. 210h/100u used for this procedure.
