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Further buffer optimization reveals that the elution can be considerably improved by increasing the conductivity through the addition of 300 mM sodium chloride, leading to a very concentrated eluate. Remarkably, merely sodium chloride at a concentration of 50 mM is proven to be sufficient for calcium-dependent antibody release in a cost-efficient manner. Antibodies of subclasses IgG2 and IgG4 are eluted with sodium chloride at neutral pH and IgG1 at pH 6, due to varying affinities for the tetrameric ZCa, ranging between 90-780 nM. The m
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