Hello Paramita

This link will be helpful.

http://www.bowdish.ca/lab/wp-content/uploads/2012/10/Protein-A-Sepharose-purification-of-Antibodies.pdf

Best.

Use Protein A&G resins as batch purification or in gravity flow column chromatography formats...

As you mentioned "specific" IgG, primarily affinity chromatography with the respective antigen can be used. This is a relatively delicate method, which might need some optimization. Could you tell us more about the purpose of this purification? It is often not necessary to purify rabbit sera if you want to use it for immunoassays.

Michael G. Weller I want to use the antibody in per-clinical study. I will tag the antibody with fluorophore and inject the same in animal to look into the interaction of protein and antibody. That is why, I want it in purified form.

Also, before affinity chromatography, to remove albumin, globulin which technique should I use:

Affinity chromatography removes all the impurities you mentioned. However, sometimes it is helpful to perform several purification steps, for example, if you need a very high purity. The first step might be Protein A.

I guess you used purified antigen to immunize Rabbit and you want to purify IgG from Rabbit serum? If so, please try mAb trap kit (commercially available). It may help your work.

Hi all.

Nice answer Michael

affinity chromatography using Protein A, G, L does not rule out self IgG from specific IgG. Immunochromatography with immobilized specific Ag is an elegant method to purify specific Ab.on the other hand, two affinity chromatography steps are not useful because use two acid elution that may lead to Ab denaturalization