What is passed over the HIC column during the first step of Hydrophobic Interaction Chromatography?

During the first step of Hydrophobic Interaction Chromatography (HIC), the supernatant containing bacterial proteins and GFP is passed over the HIC column. This is the correct approach because the HIC technique is designed to separate proteins based on their hydrophobic properties.

In the context of GFP purification, the bacterial culture is lysed to release proteins, including GFP, into the solution. The resulting supernatant contains a mixture of soluble proteins, with GFP being one of the target proteins. When this supernatant is applied to the HIC column, proteins with hydrophobic regions interact with the hydrophobic surface of the column, allowing for separation based on their hydrophobic characteristics. This step is crucial for effectively isolating GFP from other bacterial proteins, as it exploits the different hydrophobic properties of the proteins present in the supernatant.

Other options, such as sediment containing lipids or colony isolates, do not represent the composition that is typically processed during HIC. Lipids would not be targeted for purification through this method, and colony isolates are not directly used in this step, as they need to be transformed into a suitable liquid culture first. Pure GFP is the final product obtained after consecutive purification steps, and it is not passed