BIOC 2010 Lecture Notes - Lecture 12: Affinity Chromatography, Glutathione, Chelation

• School: University of Windsor
• Department: Chemistry and Biochemistry
• Course: BIOC 2010
• Professor: Michael Boffa

Document Summary

If we know the structure of a protein, or we experimentally determines its pi, we can design an ion exchange experiment to specifically isolate our protein of interest. Depending on the ph and pi, your protein will be either positively or negatively charged and will either stick or repel. Separation of different proteins based on their ability to bind specific molecules. Take insoluble matrix, but instead of a small charged groups, we attach a ligand that is specifically bound by our protein of interest. Protein sticks to column and contaminant proteins wash through. Because it is non-covalent, it is reversible. Simply apply to the column a soluble version of the ligand: it will compete with the column-bound version of the ligand for binding with the protein of interest. By adding an excess amount, you end up driving protein of interest off the column and you can collect it from the bottom.