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    Purification of Peptides

    Using High Performance Flash Chromatography

    When a peptide is synthesized and cleaved from a resin, the next step is usually a quick preparation and multiple injections onto a preparative HPLC column for isolation from deletion sequences and other contaminants. A great step to take before preparative HPLC is a single injection onto a flash purification system. This quick clean-up is often sufficient to move to the next step in the synthesis. Flash purification prior to preparative HPLC also creates the opportunity to further purify the peptide of interest with fewer injections and a less challenging mixture, and extends the life of expensive preparative HPLC columns.

     

     

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    Purification of Peptides

    High Performance Flash Chromatography (HPFC) is a powerful technique for reversed-phase purification of peptides. For twenty years, Biotage has been a pioneer in exploring new areas where the high capacity, speed and low solvent use of flash purification can be put to work. The latest and best example is simplified peptide purification with Sfär Bio columns. These columns feature 20 micron spherical particles with a large pore size of 300 Å and are a great match for the purification of peptides.

     

    Why use High Performance Flash Chromatography?

    • Enables fast and efficient purification of
      peptides and larger molecules.
    • Increased loading capacity compared to prep RP-HPLC which allows more peptide to be processed in a single injection.
    • Reduction in solvent usage and therefore
      waste compared to RP-HPLC.
    • Depending on the quality of the peptides synthesized, flash chromatography can be adopted in the workflow either as the sole method of purification or as a front end clean-up prior to RP-HPLC.
    • Our flash chromatography solutions have been used to purify peptides, PNA, peptoids and peptidomimetics.
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    Product groups

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    Biotage® Selekt

    A two column channel system with a colorful and intuitive user interface on a large touchscreen enables even the most complex tasks to become simplicity itself. High flowrate and pressure make separation faster and greener due to less solvent consumption.

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    Isolera™ Dalton 2000

    The Isolera™ Dalton 2000 system consists of Biotage® Dalton 2000 Mass Detector, Isolera™ Dalton Nanolink and Isolera™ Spektra for efficient and automated purification. Isolera Dalton 2000 directs flash fractionation by molecular mass, and in real time.

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    Biotage® Sfär Bio C4 D

    High performance flash chromatography is a very efficient technique to clean-up synthetic peptides. Biotage® Sfär Bio flash columns, with a small particle size (20 μm) and large pore size (300 Å), provide increased resolution and effective separation of complex peptide mixtures. C4 is more suitable for more hydrophobic peptides.

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    Biotage® Sfär Bio C18 D

    High performance flash chromatography is a very efficient technique to clean-up synthetic peptides. Biotage® Sfär Bio flash columns, with a small particle size (20 μm) and large pore size (300 Å), provide increased resolution and effective separation of complex peptide mixtures.

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    Resources

    The typical peptide synthesis workflow broadly involves synthesis, purification and evaporation steps to give the final pure peptide. Although the synthesis step is the most important part of the workflow, purification and evaporation issues can impact dramatically on the efficiency of your workflow and are the cause of many bottlenecks. Biotage has developed a range of tools to improve the peptide synthesis workflow, from synthesis of crude peptides through to the final purified product. These solutions make your workflow more efficient and improve the quality of peptides you synthesize. Our workflow solutions are used in pharma, biotech, CRO and academic laboratories throughout the world.

    The study of peptides, especially for therapeutic use, has grown significantly in recent years. However, producing peptides for such research is challenging, not just in the synthesis. The molecules must be visualized to allow synthetic approaches to be designed, and after synthesis the purification and isolation of peptides is not straightforward. In order to address these challenges, Biotage has developed a holistic approach to the entire peptide workflow via an automated solution, designed for dedicated peptide researchers and those new to the field.

    Peptides have garnered much attention in a variety of applications over the last few years, and interest only continues to grow. With improved synthetic strategies, peptides are now being synthesized in greater length and complexity than ever before. Purification of peptides via flash chromatography has recently been demonstrated as a viable alternative to the more standard HPLC methods currently utilized,  offering the advantage of significantly greater loading capacity while  reducing the overall purification time. Herein we present several strategies that allow for very high purity peptide samples purified by flash chromatography.

    Other steps in the workflow

    Removing Peptide Cleavage Cocktail and evaporating DMSO is no longer a problem with Biotage® V-10 Touch.

    Biotage offer a range of manual, semi-automated and fully automated systems depending on your peptide synthesis needs. Whether you require a system that makes a single peptide or multiple peptides in parallel, we have an instrument to suit your requirements.