hi

Antigen affinity column purification is the best method in antibody purification . you should prepared an affinity column by native , recombinant antigen or antigenic peptide linked in Sepharose 4B and then pass the rabbit serum for highly purified polyclonal antibody .

http://wwwuser.gwdg.de/~jgrossh/protocols/protein-purification/CNBr-Sepha.pdf

https://www.ptglab.com/media/2716/protocols-for-web_affinity-purification_v2.pdf

good luck

A protocol for plant viruses (my field) is cross–adsorption with filtered extracts from healthy tissues diluted 1:20 in serum buffer were used to remove non-specific antibodies from a poly-clonal antibody preparation.

Hi,

you can immunoprecipitate the undesired antibodies by incubating your serum with a negative bacterial lysate. Keep the antibody to antigen ratio low (1:4 to 1:10) to ensure immune complex formation. After centrifugation and recovery of the supernatant, you should have a much cleaner antibody solution.

Patrick Jack Spencer Can you please elaborate on how to decide the Antibody Antigen Ratio? do you mean the ratio of protein content of Antibody prep and the bacterial lysate. Also for centrifugation what parameters should be considered. it will be helpful If you can share some reference.

Hi again,

I couldn´t find any paper about it but if you have access to this book, that might help:

Immunoprecipitation: Purifying the Immune ComplexesCold Spring Harb Protoc; 2006; doi:10.1101/pdb.prot4536

just remember that the antibodies you want will be in the supernatant.