Antibody purification selection guide
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Antibody purification

I will purify an antibody

I will purify an antibody fragment

Both protein A and protein G have high affinity and specificity for the Fc region of antibodies. However, they differ in their ability to bind antibodies of different species and subclasses (see table). We recommend protein A for purifying human antibodies. Protein G binds a broader range of IgG from eukaryotic species and binds more classes of IgG than protein A. Therefore, we recommend our protein G-based media to purify antibodies from other species, including rat. If you are not sure whether your antibody will bind to protein A or protein G, we suggest trying rProtein A/Protein G GraviTrap, which contains gravity flow columns packed with a combination of both media.

The table below shows the relative binding strengths of antibodies from various species to protein G and protein A.

Relative binding strengths of antibodies from various species to protein G and protein A as measured in a competitive ELISA test. The amount of IgG required to give a 50% inhibition of binding of rabbit IgG conjugated with alkaline phosphatase was determined.

Species Subclass Protein G binding Protein A binding

* Purified using HiTrap IgM Purification HP columns

† Purified using HiTrap IgY Purification HP columns

++++ = strong binding

++ = medium binding

— = weak or no binding
Human IgA — variable
IgD — —
IgE — —
IgG1 ++++ ++++
IgG2 ++++ ++++
IgG3 ++++ —
IgM* — variable
IgG4 ++++ ++++
Avian egg yolk IgY† — —
Cow ++++ ++
Dog + ++
Goat ++ —
Guinea pig IgG1 ++ ++++
Hamster ++ +
Horse ++++ ++
Koala + —
Llama + —
Monkey (rhesus) ++++ ++++
Mouse IgG1 ++++ +
IgG2a ++++ ++++
IgG2 +++ +++
IgG3 +++ ++
IgM* — variable
Pig +++ +++
Rabbit +++ ++++
Rat IgG1 + —
IgG2a ++++ —
IgG2b ++ —
IgG3 ++ +
Sheep ++ +/—

Our product offering includes a range of agarose-based media with protein A, protein G, or protein L as ligand. We provide a variety of protein A- and G-based products for purifying monoclonal and polyclonal antibodies from cell culture supernatants and serum. Capto L is our medium for purifying antibody fragments that contain a kappa light chain. These fragments include single-chain fragment variable (scFv), Fab, and domain antibodies (Dab). Most of our products are available as prepacked columns for use with chromatography systems, prepacked formats for manual purification, and loose media (resins). We also offer prepacked columns for IgM and IgY purification, which are based on thiophilic adsorption.

Please use our online selection tool to find a product that meets your needs.