Leaders in monolith chromatography

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  • CIM® @Tf-0.2 monolithic 96-well plate for immunoaffinity isolation of transferrin from human plasma CIM____Tf-0_2_monolithic_96-well_plate_for_immunoaffinity_isolation_of_transferrin_from_human_plasma

    Transferrin (Tf) is a glycoprotein that transports iron to cells and has two N-glycosylation sites in humans – at asparagine 432 and asparagine 630. Carbohydrate-deficient Tf, which lacks one or both N-glycans, is the most common marker for congenital disorders of glycosylation.1 Altered Tf glycosylation has also been reported in hepatocellular carcinoma2 and chronic alcohol consumption.3,4 High-throughput Tf purification and glycan characterisation methods are under extensive development in order to facilitate screening of glycosylation patterns for population, genetic and clinical studies.

    This application note describes the development of an immunoaffinity purification method on a CIMac™ analytical column with immobilised anti-transferrin antibodies (@Tf) and the successful transfer of the method to the monolithic 96-well plate (CIM® @Tf-0.2 monolithic 96-well plate). The affinity purification method has been used for Tf isolation from human blood plasma followed by ultra-performance liquid chromatography (UPLC) analysis of Tf N-glycosylation.

  • Native elution in immunoaffinity chromatography enables highly effective virus purification Native_elution_in_immunoaffinity_chromatography_enables_highly_effective_virus_purification

    Downstream processing of viruses in virus vaccine or virus vector production accounts for up to 70% of the overall production costs. Immunoaffinity chromatography is a powerful purification technique due to its high specificity but is disadvantaged by the fact that the elution of the target molecule requires conditions for disrupting interactions between antigen and the antibody and these are often detrimental for both the immobilized proteins and target antigens, especially viruses. Here we describe the mumps virus purification using monolith-based immunoaffinity stationary phase and recently invented native elution of the bound viruses using amino acid solutions under physiological pH.

  • Increasing productivity of pDNA downstream processing using sample displacement chromatography Increasing_productivity_of_pDNA_downstream_processing_using_sample_displacement_chromatography

    Plasmid DNA (pDNA) as a pharmaceutical product has stringent requirements of purity and efficacy and often one or more chromatographic steps are used in the downstream process. High ligand density butyl-modified chromatographic monolith (CIMmultus™ C4 HLD, part of CIMmultus™ HiP² Plasmid Process Pack™ 1-1, product number 100.0011-2) is currently used in a polishing step of a pDNA purification process (1), is mainly used for separation of supercoiled (sc) pDNA separation from open circular (oc) and linear pDNA isoforms as well as for removal of remaining gDNA and RNA.
    This application note presents a comparison of two different polishing processes employing monoliths, namely bind-elute (BE) and the more recently described (2) sample displacement purification (SDP).

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