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 an alternative to reversed. Sepharose cl4b hydrophobic interaction chromatography media. Hydrophobic interaction chromatography hic is a powerful technique used for the purification of proteins in analytical. The courses are designed to provide a learning experience for process development and manufacturing staff.
Principles of hydrophobic interaction chromatography. Hic is used in the purification of proteins over a broad range of scalesin both analytical and preparatory scale. 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. Samples should be in a high salt concentration to promote hydrophobic interaction. Principles of hydrophobic interaction chromatography youtube. 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. Hydrophobicinteraction membrane chromatography hic is used to purify molecules based on differences in their hydrophobicity.
In general the conditions under which hydrophobic interaction chromatography is used are relatively mild and protein friendly resulting in. Hydrophobic interaction chromatography is a separation technique that uses the properties of hydrophobicity to separate proteins from one another. Hydrophobic interaction chromatography an overview. Hydrophobic interaction chromatography hic occupies a unique niche as a polishing step in many monoclonal antibody mab purification processes. The separation is based on the reversible interaction between a protein and the hydrophobic surface of a chromatographic medium. Hydrophobic interaction chromatography can be used for capture, intermediate purification, or polishing steps.
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. Prepacked chromatography columnsformatsreference guide. The octyl group is covalently coupled to a crosslinked 4% agarose matrix. Affinity chromatography principles and methods ge healthcare 18102229 ae 102007.
Hydrophobic interaction and reversed phase chromatography, principles and methods, 11001269 multimodal chromatography multimodal ligands capture target molecules through multiple types of interaction. Range of selectivities from low to high hydrophobicity. Chromatography resin market by type natural, synthetic. Ge healthcare hic hydrophobic interaction chromatography media features high chemical stability, properties to increase productivity and epoxy coupling manufacturer. Automated hydrophobic interaction chromatography column.
Th e protein purification handbook from ge healthcare is recommended as a. 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. Hilic uses hydrophilic stationary phases with reversedphase type. A chromatographic matrix containing hydrophobic groups, binds proteins from aqueous solutions to different extents depending on the protein structures and a range of controllable factors including concentrations of salts, ph, temperature and organic solvents fig 2. Hic offers an orthogonal selectivity to ion exchange chromatography and can be an effective step for aggregate clearance and host cell protein reduction. 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. Hydrophobic interaction chromatography hic is a widely used.
Pdf phenyl sepharose high performance butyl sepharose. This interaction is enhanced by high ionic strength buffer, which. 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. Ion exchange chromatography hydrophobic interaction and. Pdf introduction to gel filtration and hydrophobic interaction. Measured by size exclusion chromatography, specified as monomertoaggregate content. Hydrophobic interaction chromatography hic is commonly used as a polishing step in monoclonal antibody purification processes. Hydrophobic interaction chromatography ge healthcare life. In this type of chromatography, hydrophobic groups such as phenyl, octyl, or butyl, are attached to the stationary column. Hydrophobic interaction chromatography gfp purification part. 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. Sepharose cl4b hydrophobic interaction chromatography.
Hydrophobic interaction chromatography market new report by. Source 15 iso isopropyl is a highperformance resin for hydrophobic interaction chromatography designed for rapid, high resolution purification of proteins and peptides. Hydrophobic interaction chromatography hic is a valuable tool used in protein purification applications. Hydrophobic interaction chromatography ge healthcare bio. Reversedphase chromatography rpc, size exclusion chromatography sec, hydrophobic interaction chromatography hic, and affinity chromatography are often employed for assessing heterogeneity and impurity. 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. Hydrophobic interaction chromatography hic newsmedical. Pdf phenyl sepharose high performance butyl sepharose high. 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.
Ge healthcare is one of the leading players operating in the hydrophobic interaction chromatography market. Hydrophobic interaction chromatography hic is a powerful tool for the process purification of biomolecules. Ge healthcare offers a wide variety of prepacked columns and readytouse chromatography media. Hydrophobic interaction and reversed phase chromatography ge healthcare uk ltd. For many cases, hydrophobic interaction chromatography hic is an ideal separation method. In hydrophobic interaction chromatography the matrix material is lightly substituted with hydrophobic groups. Ionic interactions are commonly involved, but others such as hydrogen bonding and hydrophobic interactions can be significant. 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 absorption at 280 nm and using the extinction coefficient for the target protein. These groups can range from methyl, ethyl, propyl, octyl, or phenyl groups. Hydrophobic interaction chromatography arose mance rose high e 1. 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. Fast trak education is one means by which ge healthcare provides application training in the various aspects of bioprocessing. Wo2007064281a1 hydrophobic interaction chromatography.
Hydrophobicinteraction membrane chromatography for large. Ge healthcare hic hydrophobic interaction chromatography media features high chemical stability, properties to increase productivity and epoxy coupling. Hydrophobic interaction chromatography toyopearl hydrophobic interaction chromatography hic is a widelyused technique for separation and purification of proteins and peptides. Theory and use of hydrophobic interaction chromatography in. Chromatography principles ge healthcare life sciences.
Hydrophobic interaction chromatography cytiva, formerly. Samples are adsorbed to the resin at relatively high salt concentrations and eluted by applying. Hic, however, suffers from the limitation of use of high. The range of additives that can be used to enhance performance also complicates choices. Ge healthcare hic hydrophobic interaction chromatography media. An automated method for identifying suitable hydrophobic interaction chromatography hic media to be used in the process of protein purification is presented. The technique utilizes the accessible hydrophobic regions located on protein surfaces and their interactions with a weakly hydrophobic stationary phase.
Solute and mobile phase contributions to retention in hydrophobic interaction chromatography of proteins. This means hic is wellsuited for capture steps after sample cleanup by ammonium sulfate precipitation or for intermediate steps directly after an ion. The kit provides you with the possibility to screen for the most appropriate hic media to. Hydrophobic interaction chromatography hic process. Proteomicsprotein separations chromatographyhydrophobic. Hydrophobic interaction chromatography for purification of.
Hydrophobic interaction chromatography hic separates proteins according to differences in their surface hydrophobicity. 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. Ge healthcare hic hydrophobic interaction chromatography. 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. This video describes the principles of the technique.
It is an efficient mode to remove dimers and highmolecularweight aggregates in a monoclonal antibody purification process as a polishing step 17. Ge healthcare adopted inorganic strategies to dominate the hydrophobic interaction chromatography market. Hydrophobicity hydrophobic interaction chromatography reversed phase chromatography. 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. 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. Purification of monoclonal antibodies by hydrophobic. 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. There are handson training courses on column packing, basic chromatography, optimization and scaleup for. 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. Sep, 2012 hydrophobic interaction chromatography separates biomolecules, under relatively mild conditions, according to differences in their hydrophobicity. Hydrophobic interaction chromatography hic hic separates proteins with differences in hydrophobicity. 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.
Ge healthcare, amersham place, little chalfont, buckinghamshire, hp7 9na, uk. Substances are separated on the basis of their different hydrophobicity. Contact your local ge healthcare representative for the most current information. In my experience, hic is finding dramatically increased use both in laboratory and production processes. Instructions 71708100 ad hydrophobic interaction chromatography ge healthcare octyl sepharose cl4b octyl sepharosetm cl4b is a separation medium for hydrophobic interaction chromatography hic. The method utilizes a mediumpressure liquid chromatography system including automated buffer blending, dynamic sample loop injection, sequential column selection, multiwavelength. Ge healthcare offers a wide variety of prepacked columns, ready to use media, and pre. Ge healthcare uk ltd, amersham place, little chalfont, buckinghamshire, hp7 9na, uk ge healthcare biosciences corp. Pdf introduction to gel filtration and hydrophobic. Since the molecular mechanism of hic relies on unique.
Hydrophobic interactions between proteins and the chromatographic matrix can be exploited to purify proteins. Source iso for resin hydrophobicity requirements high resolution separations at high loads and high flow rates. Hydrophobic interaction chromatography separates biomolecules, under relatively mild conditions, according to differences in their hydrophobicity. Hitrapconvenient protein purification fisher scientific. Hic sorts biomolecules by degree of their surface hydrophobicity. A copy of these terms and conditions is available on request. Hydrophobic interaction and reversed phase chromatography. Dec 04, 2008 hydrophobic interaction chromatography.
674 703 1328 96 681 440 739 575 1257 127 1384 1525 282 960 1542 956 1428 251 45 833 223 1104 1381 672 946 191 869 617 1130 936 1495 942 1445 800 556 232 133 642 1069 1286