Hydrophobic Interaction HPLC Chromatography Services
The team at GL Technologies works with a variety of biopharmaceutical organizations both large and small with their HPLC calibration and HPLC certification requirements for their laboratories. We have worked hard to earn the trust of our clients and provide full-service HPLC calibration services according to their needs. Ensuring FDA cGMP compliance, product efficacy, lot repeatability, safety of patients, utilities and manufacturing equipment repeatability, and safety of personnel.
Hydrophobic interaction chromatography (HIC) is a powerful technique that has been widely used in the field of protein purification. It is based on the principle that nonpolar groups on the surface of a protein will interact with hydrophobic ligands on a chromatography resin in the presence of high salt concentrations. This interaction allows for selective separation of the target protein from other components in a mixture.
To speak with the experts about your Hydrophobic Interaction HPLC Calibration needs, please fill out our online form or give us a call!
The Basis of Hydrophobic Interaction HPLC Calibration
The basis of HIC lies in the fact that proteins have hydrophobic patches on their surface, which can interact with hydrophobic ligands on the surface of the chromatography resin. The hydrophobic patches on a protein are formed by the presence of nonpolar amino acids such as leucine, isoleucine, valine, phenylalanine, and tryptophan. These amino acids are typically found in the interior of a protein, but can also be exposed on the protein surface due to conformational changes or denaturation.
The chromatography resin used in HIC typically consists of hydrophobic ligands such as butyl, octyl, or phenyl groups attached to a solid support. The hydrophobic groups on the resin interact with the hydrophobic patches on the protein, resulting in the protein being bound to the resin. The strength of the interaction between the protein and the resin is influenced by the salt concentration of the buffer used in the chromatography process. At high salt concentrations, the hydrophobic interaction between the protein and the resin is weakened, allowing for elution of the protein from the column.
The HIC process involves several steps, including equilibration of the column with a buffer containing high salt concentration, loading of the protein sample onto the column, washing with a buffer containing decreasing salt concentration to remove non-specifically bound impurities, and elution of the protein with a buffer containing low salt concentration. The eluted protein can be further purified by additional chromatography steps or other purification techniques.
One of the advantages of HIC is its compatibility with a wide range of buffer conditions, including high salt concentrations and denaturants such as urea or guanidine hydrochloride. This allows for purification of proteins that are difficult to purify using other chromatography techniques. HIC is also a relatively gentle purification technique, which can help to preserve the activity and stability of the target protein.
Hydrophobic interaction chromatography is a powerful technique for the purification of proteins based on their hydrophobic properties. It is a versatile and gentle purification technique that can be used to purify a wide range of proteins under different buffer conditions.
To speak with the experts about your Hydrophobic Interaction HPLC Calibration needs, please fill out our online form or give us a call!