Antibody Purification And Immunoprecipitation

Your antibody purification and analysis workflow includes sample preparation, filtration, purification, purity check and Western blotting for protein identification and/or quantitation. Each of the steps and products selected will influence the results in terms of recovery, purity and analytical quality, but will also open opportunities to save time and money.

Use of affinity chromatography for antibody purification - how does antibody purification work?

Antibodies are members of a family of molecules, the immunoglobulins. Polyclonal antibodies, monoclonal antibodies (mAb), and antibody fragments are usually purified by affinity chromatography. Resins containing an immobilized ligand (e.g., protein A, protein G, or protein L) are used to capture antibodies and antibody fragments.

What do antibody purification schemes look like?

Antibody purification protocols typically are challenged by two factors. The first is specifically capturing as many antibodies as possible in the first step as well as controlling the degradation of the sample. The second is removing the remaining impurities and minimizing the aggregate content. Below you will find suitable protocols to choose from.

The 3-step protocol considers upscaling or process development needs. SEC is not used as a final step to remove aggregates, fragment or other impurities due to the limitations of sample volume. Instead, a combination of IEX steps is used.