The courses are designed to provide a learning experience for process development and manufacturing staff. Hydrophobic interaction chromatography for purification of. Gel filtration hydrophobic interaction ion exchange affinity reversed phase this handbook describes the role of affinity chromatography in the purification of biomolecules, the principle of the technique, the media available and how to select them. Chromatography resin market in drug discovery by type natural, synthetic, inorganic media, technique ion exchange, affinity, hydrophobic interaction, size exclusion, multimodal, and region global forecast to 2022. Hydrophobic interaction chromatography hic is a powerful tool for the process purification of biomolecules. Samples are adsorbed to the resin at relatively high salt concentrations and eluted by applying. Ge healthcare, amersham place, little chalfont, buckinghamshire, hp7 9na, uk. Fast trak education is one means by which ge healthcare provides application training in the various aspects of bioprocessing. Aug 05, 2014 hydrophobic interaction chromatography is based on the principle that hydrophobic chemicals on the resin surface will bind to hydrophobic patches on the gfp protein. Hydrophobic interaction chromatography hic separates proteins according to differences in their surface hydrophobicity. Source 15 iso isopropyl is a highperformance resin for hydrophobic interaction chromatography designed for rapid, high resolution purification of proteins and peptides.
The separation is based on the reversible interaction between a protein and the hydrophobic surface of a chromatographic medium. Ge healthcare offers a wide variety of prepacked columns and readytouse chromatography media. Phenyl sepharose cl4b instructions 71708000 ae hydrophobic interaction chromatography ge healthcare phenyl sepharose tm cl4b is a separation medium for hydrophobic interaction chromatography hic. Hydrophobic interaction chromatography hic is commonly used as a polishing step in monoclonal antibody purification processes.
Automated hydrophobic interaction chromatography column. Hydrophobic interaction chromatography hic is a valuable tool used in protein purification applications. In general the conditions under which hydrophobic interaction chromatography is used are relatively mild and protein friendly resulting in. Hydrophobic interaction chromatography hic newsmedical. Prepacked chromatography columnsformatsreference guide. This interaction is enhanced by high ionic strength buffer, which. Hitrapconvenient protein purification fisher scientific. Ge healthcare hic hydrophobic interaction chromatography media features high chemical stability, properties to increase productivity and epoxy coupling. The range of additives that can be used to enhance performance also complicates choices. Pdf introduction to gel filtration and hydrophobic. Range of selectivities from low to high hydrophobicity. Sepharose cl4b hydrophobic interaction chromatography. Measured by absorption at 280 nm and using the extinction coefficient for the target protein.
Hic offers an orthogonal selectivity to ion exchange chromatography and can be an effective step for aggregate clearance and host cell protein reduction. Hydrophobic interaction chromatography toyopearl hydrophobic interaction chromatography hic is a widelyused technique for separation and purification of proteins and peptides. Principles of hydrophobic interaction chromatography. The technique utilizes the accessible hydrophobic regions located on protein surfaces and their interactions with a weakly hydrophobic stationary phase. Hic is used in the purification of proteins over a broad range of scalesin both analytical and preparatory scale. Hydrophobic interaction chromatography hic is a commonly used technique that exploits these hydrophobic features of proteins as a basis for their separation even in complex biological mixtures. Hydrophobic interactions between proteins and the chromatographic matrix can be exploited to purify proteins. This means hic is wellsuited for capture steps after sample cleanup by ammonium sulfate precipitation or for intermediate steps directly after an ion. Theory and use of hydrophobic interaction chromatography in. Hydrophobic interaction and reversed phase chromatography. Hydrophobic interaction chromatography hic is an alternative to reversed.
For many cases, hydrophobic interaction chromatography hic is an ideal separation method. Data file 18117287 ad hydrophobic interaction chromatography phenyl sepharose high performance butyl sepharose high performance 1 hydrophobic interaction chromatography 1 media characteristics 2 laboratory and bioprocess pack sizes 2 prepacked columns 2 hiprep phenyl hp 1610 3 hiscreen phenyl hp columns 3. Hydrophobic interaction and reversed phase chromatography, principles and methods, 11001269 multimodal chromatography multimodal ligands capture target molecules through multiple types of interaction. Hydrophobic interaction and reversed phase chromatography principles and methods 11001269 2d electrophoresis using immobilized ph gradients principles and methods 80642960 microcarrier cell culture principles and methods 18114062 gst gene fusion system handbook 18115758 af. Hydrophilic interaction chromatography or hydrophilic interaction liquid chromatography, hilic is a variant of normal phase liquid chromatography that partly overlaps with other chromatographic applications such as ion chromatography and reversed phase liquid chromatography. Hydrophobic interaction chromatography is a separation technique that uses the properties of hydrophobicity to separate proteins from one another. In my experience, hic is finding dramatically increased use both in laboratory and production processes. Hydrophobic interaction chromatography hic is a commonly used technique that exploits these hydrophobic features of proteins as a basis for their separation even in complex biological mixtures 1 2. Sep, 2012 hydrophobic interaction chromatography separates biomolecules, under relatively mild conditions, according to differences in their hydrophobicity. Hydrophobicinteraction membrane chromatography for large. Substances are separated on the basis of their different hydrophobicity. Hydrophobic interaction chromatography can be used for capture, intermediate purification, or polishing steps. Ge healthcare data file 18114321 ac hydrophobic interaction chromatography hitrap hic selection kit introduction the hitrap hic selection kit consists of seven hydrophobic interaction chromatography hic media with different hydrophobic characteristics.
Reversedphase chromatography rpc, size exclusion chromatography sec, hydrophobic interaction chromatography hic, and affinity chromatography are often employed for assessing heterogeneity and impurity. The kit provides you with the possibility to screen for the most appropriate hic media to. Th e protein purification handbook from ge healthcare is recommended as a. Hydrophobic interaction chromatography hic is a reversedphase separation technique that uses a weakly hydrophobic stationary phase and a negative ionic strength gradient in a buffered aqueous mobile phase for the separation of proteins 5, 209211, 368371. Measured by size exclusion chromatography, specified as monomertoaggregate content. Ge healthcare adopted inorganic strategies to dominate the hydrophobic interaction chromatography market. Dec 04, 2008 hydrophobic interaction chromatography. These groups can range from methyl, ethyl, propyl, octyl, or phenyl groups. The method utilizes a mediumpressure liquid chromatography system including automated buffer blending, dynamic sample loop injection, sequential column selection, multiwavelength. Ge healthcare is one of the leading players operating in the hydrophobic interaction chromatography market. Principles of hydrophobic interaction chromatography youtube. In hydrophobic interaction chromatography the matrix material is lightly substituted with hydrophobic groups. In this type of chromatography, hydrophobic groups such as phenyl, octyl, or butyl, are attached to the stationary column.
Ge healthcare hic hydrophobic interaction chromatography media. Ge healthcare hic hydrophobic interaction chromatography. Principles and methods hydrophobic interaction and reversed phase chromatography principles and methods ge healthcare bioprocess, bpg, drop design, ecl, ecl plus, ettan, fineline, fplc, hiload, hiprep, hitrap, hybond, microspin, mono q, plusone, resource, sephadex. Hydrophobic interaction chromatography ge healthcare life. Interaction principle according to hydrophobic interaction and reversed phase chromatography principles and methods ge healthcare 4 close to the surface of the hydrophobic ligand and solute, the water molecules are more highly ordered than in the bulk water and appear to shield off the hydrophobic ligand and solute molecules. Wo2007064281a1 hydrophobic interaction chromatography. Since the molecular mechanism of hic relies on unique. Hydrophobic interaction chromatography hic is a powerful technique used for the purification of proteins in analytical. Today, ge healthcare offers a wide range of hic media, covering strong to weak hydrophobic interactions, on various matrices and with a range of bead sizes to fit.
Chromatography principles ge healthcare life sciences. Hydrophobicity hydrophobic interaction chromatography reversed phase chromatography. Pdf phenyl sepharose high performance butyl sepharose. Hydrophobicinteraction membrane chromatography hic is used to purify molecules based on differences in their hydrophobicity. Sepharose cl4b hydrophobic interaction chromatography media. The present invention relates to a method of isolating target compounds from a liquid, which method comprises providing a mobile phase, which comprises at least one target compound and wherein the conductivity corresponds to 0.
Proteomicsprotein separations chromatographyhydrophobic. Hydrophobic interaction chromatography arose mance rose high e 1. It is an efficient mode to remove dimers and highmolecularweight aggregates in a monoclonal antibody purification process as a polishing step 17. There are handson training courses on column packing, basic chromatography, optimization and scaleup for. Hydrophobic interaction chromatography is based on the principle that hydrophobic chemicals on the resin surface will bind to hydrophobic patches on. Hydrophobic interaction chromatography eriksson 2011. Data file 29031925 ab hydrophobic interaction chromatography ge healthcare life sciences capto phenyl impres and capto butyl impres capto phenyl impres and capto butyl impres are hydrophobic interaction chromatography hic media resins developed for the intermediate and polishing steps in a downstream protein purification process fig 1. An automated method for identifying suitable hydrophobic interaction chromatography hic media to be used in the process of protein purification is presented. Hydrophobic interaction and reversed phase chromatography ge healthcare uk ltd. Hydrophobic interaction chromatography cytiva, formerly. Hydrophobic interaction chromatography an overview. Chromatography resin market by type natural, synthetic.
Today, ge healthcare offers a wide range of hic media, covering strong to weak hydrophobic interactions, on various matrices and with a range of bead sizes to fit most applications and scales. Source iso for resin hydrophobicity requirements high resolution separations at high loads and high flow rates. Ionic interactions are commonly involved, but others such as hydrogen bonding and hydrophobic interactions can be significant. Ge healthcare uk ltd, amersham place, little chalfont, buckinghamshire, hp7 9na, uk ge healthcare biosciences corp. Hydrophobic interaction chromatography hic occupies a unique niche as a polishing step in many monoclonal antibody mab purification processes. Affinity chromatography principles and methods ge healthcare 18102229 ae 102007. Solute and mobile phase contributions to retention in hydrophobic interaction chromatography of proteins. The octyl group is covalently coupled to a crosslinked 4% agarose matrix. Hydrophobic interaction chromatography ge healthcare bio. Hydrophobic interaction chromatography market new report by. Hilic uses hydrophilic stationary phases with reversedphase type. Sepharose cl4b hydrophobic interaction chromatography media, ge healthcare protein purification media hydrophobic interaction chromatography is a versatile method for the purification and separation of biomolecules based on differences in their surface hydrophobicity. Pdf phenyl sepharose high performance butyl sepharose high. Hic sorts biomolecules by degree of their surface hydrophobicity.
Samples should be in a high salt concentration to promote hydrophobic interaction. Hydrophobic interaction chromatography separates biomolecules, under relatively mild conditions, according to differences in their hydrophobicity. Ge healthcare offers a wide variety of prepacked columns, ready to use media, and pre. This video describes the principles of the technique. Purification of monoclonal antibodies by hydrophobic. Hydrophobic interaction chromatography gfp purification part. Ion exchange chromatography hydrophobic interaction and. Hic, however, suffers from the limitation of use of high.
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