I have raised an Antibody against BSA-hapten conjugate. Purified the antibodies with Protein A/G column affinity chromatography. Now, I have a mixture of antibodies consisting of anti-BSA Ab and anti-BSA-hapten conjugate Ab. I want to purify anti-BSA-hapten conjugate Ab from anti-BSA Ab via passing through BSA coupled SiO2 gel column. My hypothesis is that anti-BSA Ab will bind to the BSA molecule attached to the SiO2 bead and the remaining anti-conjugate Ab will come out. Since, I need only anti-conjugate Ab, therefore, I don't need to elute out anti-BSA Ab. But, here I would like to know about the method to cross-link BSA with SiO2 beads. Kindly, suggest.
I don't know how to couple proteins to silica, but I would recommend instead using cyanogen bromide-activated Sepharose, which is a good resin for protein compatibility, and it is a very simple procedure.
https://www.sigmaaldrich.com/catalog/product/sigma/c9142?lang=en®ion=US
You can find in the following paper a method for attaching a protein to this resin. The method is for an antibody, but it works for any protein.
http://www.jbc.org/content/269/5/3745.long
You can use aptes to introduce amino groups on the surface of silica. Afterwards glutaraldehyde can be used to attach bsa along with reduction.
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