Regina Römling

Antibody related biopharmaceuticals represent one of the most dynamic segments of the pharmaceutical market. The analysis and monitoring of product titers in CHO cell cultures is one of the key tasks during development of new products. Cell line selection, optimization of expression rates, and process control need fast and efficient antibody quantification. Besides ELISA assays with known limitations with regard to reproducibility and precision, high performance liquid chromatography (HPLC) analysis using affinity chromatography is applied. This application note summarizes some performance data of a new Protein A affinity column for mAb titer analysis.

Tosoh Bioscience

In recent years water-soluble polymers are gaining more and more interest in different applications. The molecular weight distribution of polymers is usually characterized by size exclusion chromatography (SEC) coupled with refractive index, viscometric or laser light scattering detection. Recent advances in SEC comprise semi-micro SEC and the design of linear columns providing wide molecular weight separation ranges and near-linear calibrations. We describe the separation of water-soluble polymers with a new generation of linear, polymer-based, semi-micro SEC columns using the compact EcoSEC SEC system.

There is considerable attention placed on the synthesis and characterization of polypeptoids, a new class of synthetic polypeptide analogues. Because of intense hydrogen-bonding among chains of these polymers, aggressive solvents such as hexafluoroisopropanol (HFIP) must be used as the mobile phase to support solubility.

Development and production of biopharmaceuticals is a growing segment of the pharmaceutical industry. The development of vaccines is only one - but as a result of the current H1N1 flu hysteria it is the most striking - segment of this emerging field. Recombinant proteins, ranging from insulin products to therapeutic antibodies, represent another important group of biopharmaceuticals. With the introduction of the first so-called biosimilars in Europe the demand for highly efficient analysis methods is further increasing. Newly developed stationary phases for the typical modes of biochromatography of native proteins such as SEC or IEC open up new opportunities for increasing throughput in the bioanalytical lab.

Hydrophilic interaction liquid chromatography (HILIC) offers unique advantages for the separation of very polar compounds when compared to reversed-phase chromatography. A new silica based HILIC phase was developed to provide additional selectivity options in HILIC separations. The separation of water soluble vitamins on the new TSKgel NH2-100 HILIC column and on the well known TSKgel Amide-80 HILIC column demonstrates the differences in selectivity.