Is HIC a type of affinity chromatography?
Hydrophobic interaction chromatography (HIC) separates molecules based on their hydrophobicity. Related Topics: Size Exclusion Chromatography, Ion Exchange Chromatography, Mixed-Mode Chromatography, Affinity Chromatography, Low Pressure Chromatography Systems and Medium Pressure Chromatography Systems.
How does HIC work?
Hydrophobic interaction chromatography (HIC) separates proteins according to differences in their surface hydrophobicity. HIC utilizes a reversible interaction between the proteins and the hydrophobic ligand of a HIC resin. Lowering the salt concentration weakens the interaction.
Why does high salt favor hydrophobic interactions?
The salt promotes interaction between the hydrophilic and hydrophobic regions of the protein and the medium by reducing the solvation of sample molecules and exposing their hydrophobic regions. Hence the sample molecules can be eluted out in the order of increasing hydrophobicity using a decreasing salt gradient.
What is HIC column?
In a nutshell, HIC (also known as ‘salting out’) separates protein molecules using the properties of hydrophobicity. In this method, proteins containing both hydrophilic and hydrophobic regions are applied to an HIC column under high salt buffer conditions.
What is HIC and why is it appropriate for purification of GFP?
Unique characteristics of GFP enable it to be purified from bacterial cell proteins using HIC columns. 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 through the column.
Does salting out cause protein denaturation?
The starting molecules strengthen hydrophobic interactions by decreasing solubility of the nonpolar molecules, thus salting out the system. However, the later molecules begin to denature the structure of the protein because of strong ionic interactions that disrupt hydrogen bonding.
What is hydroxyapatite and how and where is it made?
Hydroxyapatite is a naturally occurring form of the mineral calcium apatite—calcium, phosphorous, and oxygen—that grows in hexagonal crystals. Pure hydroxyapatite is white in color. It makes up most of the human bone structure, builds tooth enamel, and collects in tiny amounts in part of the brain.
How do you separate proteins from salt?
At a very high ionic strength, protein solubility decreases as ionic strength increases in the process known as ‘salting-out’. Thus, salting out can be used to separate proteins based on their solubility in the presence of a high concentration of salt.