GFP Chromatography
Chromatography 101. Chromatography to purify proteins of interest depends on a protein's chemical or physical properties, such as molecular weight, electrical charge, or solubility. Green Fluorescent Protein (GFP) is extremely hydrophobic compared to bacterial proteins. This unique characteristic of GFP enables the purification of GFP from bacterial cell proteins using hydrophobic interaction chromatography (HIC). When placed in a buffer containing a high concentration of salt, the HIC matrix selectively binds hydrophobic GFP molecules while allowing the bacterial proteins to pass right through the column. Then, simply lowering the salt concentration of the buffer causes GFP to elute from the column in pure form.
pGLO Bacterial Transformation plate and a Series of HIC columns showing the separation of GFP.