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Tosoh Bioscience - Your Specialist in Separation White Papers

Tosoh Bioscience - Your Specialist in Separation

Smart HPLC Solutions for Large Molecules The biopharmaceutical research sector and its products represent a significant area of growth for today’s pharmaceutical industry. Major biotherapeutic categories currently include vaccines, therapeutic proteins such as growth factors or cytokines, and, last but not least, monoclonal antibodies. A thorough characterisation of these products is a key task for the successful submission of data for their regulatory approval.
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TOYOPEARL gigaCap® Series ion exchange Chromatography (iEC) is one of the most frequently used chromatographic modes for the separation and purifi cation of biomolecules. it is used at all stages and scales of purifi cation of therapeutic proteins: from laboratory scale purifi cation to industrial scale downstream processing (dSP). toyopearl media are hydrophilic, macroporous polymethacrylate resins, available with various ligands and in different particle and pore sizes.
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TOyOPEARl® AF-rProtein A-650F Protein A based affi nity chromatography is the industry standard for the capture of monoclonal antibodies (MAbs) in downstream processing. Affi nity media offer high selectivity thereby yielding a high purity of the target molecule in one step. The antibodies are captured at near neutral pH and eluted using acidic pH. The rising fermentation titres in the production of monoclonal antibodies drive the focus on developing resins that increase downstream capacity and effi ciency for these processes.
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Direct analysis of biogenic amines in food by HILIC-MS Biogenic amines are present in protein-rich food such as fish, meat or milk. Their concentration increases during food decomposition. The reasons to monitor biogenic amines in food products are to determine that the food is suitable for consumption and to establish appropriate storage conditions.
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Efficient SEC-UV-RI-MALS Analysis of Protein Aggregates Static light scattering in combination with size exclusion chromatography is a valuable tool for verifying purity of monoclonal antibodies (MABS) as such or as a quick check while the downstream processing takes place. Besides fluorescence detection, light scattering is one of the most sensitive methods to detect protein aggregates.
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Analysis of Saccharides by HILIC using TSKgel® NH2-100 Columns Saccharides are fundametal substances that express various bioactivities and may exist independently or form complexes with proteins or lipids. They can be classified into monosaccharides, disaccharides, oligosaccharides, polysaccharides, based upon the degrees of polymerization and condensation.
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Analysis of Water-Soluble Polymers Using Linear Size Exclusion HPLC Columns and a Semi-Micro SEC System 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.
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CHARACTERIZING BIOTHERAPEUTICS WITH HPLC Development and production of biopharmaceuticals is a growing segment of pharmaceutical industry. Therapeutic proteins include blood factors, thrombolytic agents, growth factors, interferons, interleukins, therapeutic enzymes and last but not least monoclonal antibodies.
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Reversed-Phase Analysis of Alkylbenzene Sulfonates Linear alkylbenzene sulfonates (LASs) are synthetic surfactants, which are produced by alkylation and sulfonation of benzene. LASs are a mixture of homologues and phenyl positional isomers, each containing an aromatic ring sulfonated at the para position and attached to a linear alkyl chain. The product is used in detergents and cleaning products for domestic and industrial use.
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Characterisation Studies of PEGylated Lysozyme PEGylation, the process by which polyethylene glycol (PEG) chains are attached to protein and peptide drugs is a common practice in the development of biopharmaceuticals to prolong serum half-life and improve pharmacokinetics of a drug. There is increasing demand for chromatographic methods to separate the modified isoforms from the native protein.
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Glycosylation analysis by hydrophilic interaction chromatography (HILIC) – N-Glyco mapping of the ZP-domain of murine TGFR-3 Glycosylation is one of the most common forms of post-translational modification of eukaryotic proteins. Glycosylated proteins (glycoproteins) make up the majority of human proteins. The polysaccharide side chains (Glycans) play critical roles in physiological and pathological reactions ranging from immunity to cell signaling, development and death.
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Alternate Selectivity for Polar Compounds in Hydrophilic Interaction Liquid Chromatography (HILIC) Using a New Amino Type HILIC Column 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.
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Analysis of Melamine and Cyanuric Acid in Milk by LC-MS/MS Melamine is an organic base and a trimer of cyanamide, with a 1,3,5-triazine skeleton. Melamine is combined with formaldehyde to produce melamine resin, a very durable thermosetting plastic, and melamine foam, a polymeric cleaning product. The end products include countertops, dry erase boards, fabrics, glues, house wares and flame retardants. Melamine is one of the major components in Pigment Yellow 150, a colorant in inks and plastics.
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HPLC Analysis of lysozyme in different types of wine Hen’s egg white lysozyme is used in winemaking to eliminate lactic acid bacteria and to control malolactic fermentation. As eggs and derivatives are regarded as potential allergenics the European Commission issued a directive that wine labels must include information about egg derived ingredients like lysozyme. As a consequence the International Organisation for Wine and Vine (OIV) published a standard method for measurement of lysozyme in wine by HPLC, which is based on a method developed at the University of Bologna.
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Analysis of cationic polymers by Size Exclusion Chromatography with TSK-GEL® In recent years water soluble cationic polymers are gaining more and more interest in different applications ranging from hair conditioning to gene transfection and drug delivery. The molecular weight distribution of polymers is usually characterized by SEC coupled with refractive index, viscometric or laser light scattering detection.
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HILIC–MS - High Resolution and Sensitivity for the Analysis of Very Polar Compounds Hydrophilic interaction liquid chromatography (HILIC) offers unique advantages for mass spectrometric detection of very polar compounds when compared to reversed phase chromatography. The higher organic content of the eluent in HILIC supports efficient evaporation of the solvent, thus enhancing sensitivity and altering ion suppression.
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