TOYOPEARL polymeric AFC resins are available in a variety of group specific ligands. In addition, activated and reactive forms are available for the immobilization of target specific ligands. In AFC the mechanism of binding and elution varies depending on the immobilized ligand used. In some cases such as with TOYOPEARL AF-Chelate-650M (for Immobilized Metal Affinity Chromatography or IMAC) the binding and elution mechanism may change through the use of different metals or chemical modifiers. DBC values for AFC materials vary with ligand and are typically on the low side. However, because AFC has high selectivity and most of the capacity is used for the target, with feedstock impurities remaining unbound, AFC resins are typically used for the Capture and Intermediate purification steps of a manufacturing process.
Select an Affinity Resin
Protein A chromatography has become a widely used platform in monoclonal antibody (mAb) purification. It makes use of the specific interactions that take place between the Fc regions of immunoglobulin G and immobilized Protein A, a cell wall component of Staphylococcus aureus. Its high specificity typically generates more than 95% purity in one step.
Two versions of TOYOPEARL Protein A media are available, both carrying recombinant alkali stable Protein A ligands. The new TOYOPEARL AF-rProtein A HC-650F benefits from superior mAb capacity over a broad range of feedstock concentrations and residence times.
Select a Protein A Resin
Protein L affinity chromatography makes use of the specific interactions that take place between the variable region of an antibody’s kappa light chain and immobilized Protein L (Peptostreptococcus magnus). Protein L binds a wider range of antibody classes than Protein A. Typical targets are antigen binding fragments (Fabs), single-chain variable fragments (scFvs) and domain antibodies (dAbs).
The new TOYOPEARLProtein L resin is carrying a recombinant, fairly alkali stable Protein L ligand. TOYOPEARL AF-rProtein L-650F shows twice the binding capacity of other Protein L media.
Select a Protein L Resin
Hydrophobic Interaction Chromatography (HIC) is a widely used technique for separation and purification of proteins and peptides. It sorts biomolecules by degree of their surface hydrophobicity. Samples are adsorbed to the resin at relatively high salt concentrations and eluted by applying a decreasing salt gradient. The mild conditions used in HIC separation of peptides and proteins typically maintain protein structure and biologic activity.
Tosoh Bioscience offers seven product lines of TOYOPEARL HIC resins using five different ligands. The different degrees of hydrophobicity and selectivity support the user in selecting the best solution for a given target.
Select a Hydrophobic Interaction Chromatography Resin
Ion Exchange Chromatography (IEC or IEX) is the most common liquid chromatographic method used in manufacturing therapeutic proteins. Compared with other chromatographic modes, modern ion exchange media offer high dynamic binding capacities and a straightforward method development. IEX is used at all stages and scales of purification of therapeutic proteins: from laboratory scale purification to industrial scale downstream processing (DSP).
Tosoh Bioscience offers a broad range of resins for ion exchange applications. The TOYOPEARL GigaCap series of high capacity ion exchange media was developed to meet the needs in high throughput purification of monoclonal antibodies and other proteins.
Select an Ion Exchange Resin
Mixed-Mode Chromatography resins have both ionic and hydrophobic groups in their composition. These unique ligands allow the separation of acidic, basic, and neutral proteins by one resin. Unlike with traditional ion exchange resins, elevated conductivity feedstocks and buffers can be used without diminishing binding capacity.
TOYOPEARL MX-Trp-650M is a multimodal cation exchange resin with unique selectivity and high recovery. It provides the highest binding capacity among all commercially available mixed-mode resins and it offers outstanding selectivity for mAbs and mAb aggregates. Method development for mixed-mode chromatography using the tryptophan ligand is usually no more elaborate than for traditional HIC or IEX resins.
About the Mixed-Mode Resin
Size Exclusion Chromatography (SEC), also known as gel filtration when applied in an aqueous mobile phase, separates molecules based on differences in size. As a non-binding chromatographic mode, SEC does not provide binding capacities known from adsorptive modes such as IEX. At manufacturing scale, the main applications of SEC are desalting of the feedstock, removal of aggregates in a final polishing step and virus purification.
TOYOPEARL polymeric SEC materials are available in a number of pore sizes and particle diameters. It is important to understand the pore sizes, particle sizes, and physical properties of these beads, as they are the base beads for TOYOPEARL IEX, HIC, Mixed Mode and AFC products.
Select a Size Exclusion Resin
Hydroxyapatite (HA) has become one of the best characterized multimodal chromatography packings, with valuable research and industrial applications throughout the field of bioprocessing. Its combination of ion exchange and metal affinity have given it the ability to achieve separations that no other chromatography media can match, and it continues to provide unique solutions for emerging challenges in the field.
Ca⁺⁺Pure-HA provides exceptional separation properties and unequalled selectivity and resolution for multiple classes of biomolecules. The robust nature of Ca⁺⁺Pure-HA offers the flexibility to use this resin at any stage in a process from capture to final polishing.
About the Hydroxyapatite Ca++Pure-HA