Applications & Products

Other Proteins..

Mimetic Ligand^TM Technology

Mimetic Ligands^TM are highly stable, fully synthetic affinity ligands which are capable of capturing, purifying and removing a wide range of proteins. A diverse range of Mimetic Ligand^TM structures have been developed, each of which incorporates features which mimic the protein-binding components of natural compounds. Consequently each Mimetic Ligand^TM has a very different selectivity allowing a wide range of proteins to be bound and purified. Mimetic Ligands^TM are the ideal starting point for development of affinity capture steps, especially in situations where an established affinity purification platform is unavailable for your protein of interest. The current range of general-purpose Mimetic Ligands^TM includes Mimetic Red® 2, Mimetic Red® 3, Mimetic Orange® 1, Mimetic Orange® 2, Mimetic Orange® 3, Mimetic Yellow® 1, Mimetic Yellow® 2, Mimetic Green® 1, Mimetic Blue® 1 and Mimetic Blue® SA.

Unlike relatively crude textile dye preparations used previously for protein purification applications, Mimetic Ligands^TM have purpose-designed structures which are synthesised to high purities and immobilised to solid supports by extremely stable spacer-arm linkages. They offer higher degrees of purification, substantially lower ligand leakage and more reproducible separations than can be achieved with conventional textile dye adsorbents. Furthermore, Mimetic Ligand^TM adsorbents are fully resistant to treatment with sodium hydroxide and are manufactured by ProMetic BioSciences Ltd in bulk quantities to Good Manufacturing Practice standards (ISO9001).

Selection of the most appropriate Mimetic Ligand^TM for any given protein could not be simpler with the availability of Mimetic Ligand^TM screening kits. These comprise PIKSI® kits which contain 10 x 1ml columns of each Mimetic Ligand^TM adsorbent; pre-packed 1 ml column kits and a 96-column-block format Puraplate^TM which comprise 8 x 0.25ml columns of each Mimetic Ligand^TM adsorbent. Puraplates may be used manually or in conjunction with automated liquid handling robots for rapid adsorbent selection and method development.

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Hydrophobic Interaction Chromatography

Hydrophobic interaction chromatography (HIC) is an established technique for the purification of proteins based upon differences in relative hydrophobicity. PBL offers a comprehensive range of adsorbents for use in HIC applications. Different proteins have different hydrophobicities and adsorption to HIC supports can be controlled by the addition of water structuring salts such as ammonium sulphate and/or changing the surface hydrophobicity of the chromatography adsorbent. Addition and removal of salt is the commonest way of achieving binding and elution, though there is a limit to how much salt can be added without causing precipitation problems. For this reason, PBL offers a series of HIC adsorbents of increasing hydrophobicity: Butyl Agarose 6XL, Hexyl Agarose 6XL, Octyl Agarose 6XL, Phenyl Agarose 6XL and Decyl Agarose 6XL. This enables selection of a HIC adsorbent with the correct hydrophobicity for the protein to be purified and avoids the need for excessive amounts of salt for protein binding, or the use of organic co-solvents to assist with elution. PBL's range of HIC adsorbents are produced using a cross-linked agarose support matrix and epoxy-bonded HIC groups which provides robust and alkali resistant adsorbents which may be used for a wide range of protein purification applications. Selection of the most appropriate PBL HIC adsorbent is made easier with the Hydrophobic A6XL Screening Column Kit, a kit comprising of pre-packed 1 ml columns - containing each resin type. Similarly, method development is also made easier with the Hydrophobic A6XL Column Kit. These individual kits comprise of 5 x pre-packed 1ml columns - containing the same resin type. Also, the 96-column-block format Puraplate^TM is also available (each plate containing a single HIC resin type).

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Ion Exchange Chromatography

Ion exchange chromatography is used to separate proteins on the basis of their net charge at a particular pH. This technique is used extensively in the biotechnology industry and is particularly useful when used as a polishing step following an affinity capture column. Anion exchange adsorbents are used when the protein(s) to be bound are negatively charged at the selected pH, whereas cation exchangers are used when the protein(s) to be bound are positively charged. Typically, binding is performed in a low salt buffer and elution is achieved by addition of salt and/or adjustment of pH.PBL currently offers one weak cation exchanger (DEAE Purabead® HF) and one strong anion exchanger (SP Purabead® HF) with Q and CM adsorbents currently in development (please enquire for details). All of PBL's ion-exchange products are based upon Purabead® HF, a near-monodisperse high flow agarose support matrix developed specifically for bioprocess applications. The uniform particle size and excellent bead strength of Purabead® HF enables the operation of large packed bed columns and high linear flow rates.

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Specific Applications:

Protease Purification

p-Aminobenzamidine Agarose 6XL can be used to purifiy a wide variety of serine proteases and esterases. The p-aminobenzamidine ligand has specific affinity for the active site of serine proteases such as trypsin, thrombin urokinase and kallikrein. p-Aminobenzamidine Agarose 6XL from PBL has been optimised for these applications and is used commercially for the manufacture of serine protease therapeutics.

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Alkaline Phosphatase Purification

Mimetic Blue® AP has been developed specifically for the purification of alkaline phosphatase from a variety of sources including bovine, human, microbial and genetically engineered forms. This affinity adsorbent provides high levels of purification and large increases in enzyme specific activity are possible.

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Capture of biotinylated proteins

Biotinylated proteins may be captured and immobilised using Avidin Agarose 4XL. The high affinity of the biotin-avidin binding interaction means biotinylated compounds are bound extremely tightly and are effectively immobilised to the avidin agarose support. Avidin Agarose 4XL comprises hens egg avidin attached to a wide-pore 4% beaded agarose support.

For further information, please contact us at sales@prometicbiosciences.com