What is HIC used for?
Hydrophobic interaction chromatography (HIC) is a valuable tool used in protein purification applications. HIC is used in the purification of proteins over a broad range of scales—in both analytical and preparatory scale applications.
How does HIC chromatography 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.
Is HIC affinity chromatography?
HIC separates proteins and other biomolecules from a crude lysate based on differences in hydrophobicity. Similar to affinity chromatography (AC) and ion exchange chromatography (IEX), HIC is capable of concentrating the protein of interest as it progresses through the chromatographic process.
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.
What is hydroxyapatite chromatography?
Hydroxyapatite (HA) chromatography (HAC) is one of the oldest chromatography methods in the field of biomolecule separations [6, 7], and has been used for separation of proteins and DNAs [8-25]. It is regarded as a mixed-mode or multi-modal chromatography. This is similar to cation exchange chromatography (CIEC).
What is HIC and how does it function to purify proteins?
Hydrophobic interaction chromatography (HIC) separates molecules based on their hydrophobicity. HIC is a useful separation technique for purifying proteins while maintaining biological activity due to the use of conditions and matrices that operate under less denaturing conditions.
How did the HIC columns purify the protein?
What is elution buffer in HIC?
In HIC, proteins are applied to the column in a high-salt buffer and eluted in a low-salt buffer. In some experiments, researchers gradually decrease the salt concentration of the solvent, causing different proteins to elute at different rates (least hydrophobic proteins first).
What is the function of hydroxyapatite?
Hydroxyapatite is chemically similar to the mineral component of bone and hard tissues in mammals. It is a bioactive material that will support bone ingrowth and osseointegration. It has been used in orthopedic, dental, and maxillofacial applications and can be replaced with bone formation via osteoconduction.
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 did ammonium sulfate affect the HIC?
How Does HIC Work? In a nutshell, HIC (also known as ‘salting out’) separates protein molecules using the properties of hydrophobicity. The salt in the buffer (usually ammonium sulfate) reduces the solvation of sample solutes and exposes the hydrophobic regions along the surface of the protein molecule.
What is a HIC column?
Protein-Pak Hi Res HIC Columns contain non-porous, polymethacrylate-based particles (2.5 μm) functionalized with a butyl ligand coating and are well suited for the characterization of proteins and biotherapeutics including monoclonal antibodies (mAb) and antibody drug conjugates (ADC).