![]() In many applications, the cation-exchange method has shown better ability to remove host cell proteins, desquamation protein A and high molecular weight monoclonal antibodies than the anion-exchange method, in the current production of the therapeutic monoclonal antibody, macroporous polymer matrix strong cation exchange chromatography resin has been widely and successfully applied. This characteristic requires that the cation exchange chromatography resin must have excellent selectivity and dynamic loading, these two become the most important index to measure this kind of resin. As mentioned earlier, the IEP of therapeutic MAbs is usually high, so the use of cation exchange in the McAb refining process is in the binding mode rather than the flow-through mode. Methacrylate resin with polymeric “tentacles”Ĭoated cross-linked poly (styrene-divinylbenzene)Ĭoated mono dispersed Cross-linked poly (styrene-divinylbenzene)Ĭation exchange (CEX) is also one of the most common methods for the purification of McAb. Highly cross-linked agarose with dextran surface extenderĦ% cross-linked agarose with dextran surface extender Quaternary ammonium with aromatic structure Main Parameters Of Anion-exchange Chromatography Resins Products The following table shows the parameters of some anion exchange chromatographic resins. It has been reported in the literature that this method can be used as a refining method to improve the flow-through mode of anion exchange chromatography for removing the defects of the McAb molecular polymer and the protein a molecule, at the same time, it maintains the advantage of flow-through mode to remove DNA, host cell protein and endotoxin. , the target monoclonal antibody is eluted first and the impurity molecules are eluted later in the isoelution. Because the target monoclonal antibody has a higher isoelectric point than all the impurity molecules, the impurity molecules bind more strongly on the anion exchange chromatography column, i. In this method, the target monoclonal antibody (McAb) was firstly bound to anion-exchange chromatographic column with suitable pH buffer, and then the McAb was equally eluted by salt solution with certain conductivity. Weak-partitioning chromatography (WPC) is a new method to refine McAb with a combination mode. For McAb that can maintain stability at high Ph, the combination mode may be considered. In the flow-through mode, the DNA and host cell proteins have been largely removed by the protein A affinity chromatography step, which requires the anion exchange chromatography packing to have a strong adsorption capacity for these impurity molecules, but it doesn't require a very high dynamic load. The anion exchange method can also remove virus in flow-through mode, but its disadvantage is that it is not effective to remove McAb and protein A. However, too high pH has the potential to cause the monoclonal antibody molecules to mutate, so the binding mode is used less often than the flow-through mode. If a buffer with a higher pH is used, the monoclonal antibody can carry enough negative charge to remain on the anion-exchange column, known as the binding mode. Impurities, such as DNA, host cell proteins and Endotoxin, are negatively charged in this Ph range and interact strongly with the anion-exchange packings to remain on the column. The monoclonal antibody molecules do not interact with the anion-exchange chromatographic packings in any way and are often referred to as the "fluid-through mode". ![]() Ion-exchange Chromatography Menu ToggleĪnion exchange (AEX) is one of the most common methods for the purification of McAb.īecause the isoelectric point of therapeutic monoclonal antibody is usually high, the buffer solution with pH of 7 ~ 7.5 is suitable. ![]()
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