We are purifying custom-made antibodies from frozen chicken serum, and when thawed it turned into the gelatin-like substance on ice. So, their erythrocytes are nucleated and then I treated serum with DNAse, thinking that it is a genomic DNA released, it helped a little, I centrifuged, took out super, put on ice and it turned into the gelatin again. It can't be loaded on the column like that. What is it in their blood and how to get rid of it? Thank you!
Have you tried diluting the serum and possibly adding EDTA? Does it happen with fresh blood, too?
You also might try delipidating it with hexanes.
Thank you, Wolfgang! I diluted 2-fold with PBS, it did do much. I did not try EDTA, I'll try. I don't have fresh blood, they send us frozen serum. Can lipids cause gel formation? I know they do delipidation for purification from egg yolks, not from serum. I'll try.
Did you remove albumin? Pass diluted serum through an albumin depletion resin and observe the supernatant...Preparing a highly diluted sample and ultracentrifugation to overcome viscosity following the application to the immunoaffinity column for custom-made antibody purification may also work without an albumin removal strategy as mentioned earlier ...
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