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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.

  • A method for concentration and purification of human coronavirus HCoV-OC43 using CIM QA monolithic columns A_method_for_concentration_and_purification_of_human_coronavirus_HCoV-OC43_using_CIM_QA_monolithic_columns

    Human coronavirus OC43 (HCoV-OC43) is a frequent cause of respiratory tract illness, ranging from common cold to severe disease. The research on coronaviruses and medical application of coronaviral vectors/vaccines requires a quality material of high purity. Unfortunately, virus preparations are highly contaminated with cell debris and purification requires laborious, cost-ineffective procedures.
    Here, we report a simple and efficient method for coronavirus concentration and purification by the example of HCoV-OC43. To achieve this, virus chromatography was performed on CIM QA monolithic columns (BIA Separations), with immobilized positively charged quaternary amines. The quality of the obtained virus stock was assessed with SDS Page electrophoresis, followed by Western blot analysis. Finally, infectivity of recovered virus was evaluated by titration.

  • Comparison of hydrophobic monolithic supports for sample displacement chromatography of plasmid DNA Comparison_of_hydrophobic_monolithic_supports_for_sample_displacement_chromatography_of_plasmid_DNA

    Sample displacement chromatography exploits the different relative binding affinities of components in a sample mixture to achieve accummulation of a desired substance on the column before elution. In pharmaceutical applications, requirements for purity and efficacy of plasmid DNA (pDNA) as a therapeutic product are stringent. The separation of linear, supercoiled (sc) and open-circular (oc) pDNA isoforms has already been established on CIM® butyl (C4 HLD) monolithic columns at preprative scale. This process requires high concentration of ammonium sulphate for loading which increases the overall production requirements. Competing adsorption in sample displacement chromatography utilises the binding capacity of the chromatographic resin more efficiently and increases productivity of the chromatographic step.
    This application note investigates three monolithic chromatographic supports with different hydrophobicities regarding their applicability for sample displacement of pDNA. CIMac™ C4 HLD (butyl, high ligand density) as a commercial product and pyridine and histamine as custom immobilised columns are compared.

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