I'd like to bind antibodies of species 1 to a protein A (or protein G) resin and to catch antibodies of species 2 that are cross reactive against species 1. At the end I have a sandwich of Resin Ab of species 1 Ab of species 2.
I'm interested in the flow-through, which is not cross-reactive to species 1 to obtain affinity purified antibodies. Afterwards I'd like to recover the resin to use it again for the same purpose. And this is the main point of my question:
Is it possible to elute the cross-reactive antibodies of species 2 from the antibodies of species 1 without elution or destruction of the antibodies of species 1 that are directly bound to the resin, resulting in Resin Ab of species 1 + eluted/destructed Ab of species 2?
It could be possible, but Ab 1 have to be covalently conjugated with the resin. Then you can use a suitable buffer with pH that facilitates detachment of Ab 1 from Ab 2. Covalent conjugation of Ab 1 to the resin could be made by using a carboxyl resin, activated with water-soluble carbodiimide. Similar conjugations are used for example in SPR analysis of antibody affinity to antigens.
Just a short update - and a new question. I came up working with the AminoLinkPlus-Kit (Thermo Scientific) which works pretty good for my purpose. The only thing is the elution which is not complete using 0,1M Glycin, pH 2,8. I need to split an antibody-antibody complex, keeping both antibodies more or less intact. Is there any trick like adding high salt concentrations or other additives?
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