The optimal purification method is dictated by the species, the source material, and the level of immunoglobulin purity required. The amount of supernatant or serum required per column depends on the concentration and total milligrams of antibody in the solution.
Antigen affinity
If a high purity of antibody is required, a
target-specific peptide or
recombinant protein affinity column can be used to obtain extremely pure antibodies. A minimum of 10 mg of
pure antigen (or purified peptide) is required for the generation of custom resins to capture the target
protein. Due to the highly specific purification process, the resulting antibody preparationtends to have
lower background levels and lower non-specific binding rates than other purification methods.
Protein A and Protein G
Protein A and Protein G are the most common
choices for antibody
purification due to their ability to bind the constant (Fc) region of IgG from various species. Most intact
IgG antibodies can be purified efficiently by Protein A or Protein G, but their affinity differs by species
and subclass of IgG.
Aseptic/Low Endotoxin Purification
Purification is conducted under
aseptic conditions in a
dedicated, limited-access facility to generate clean, low-endotoxin antibody preparations.
Ammonium sulfate precipitation
Ammonium sulfate is widely used in
isolating both IgY and IgG from
a variety of species. An antigen affinity purification step can be employed to further purify the
immunoglobulin of interest.
IgM purification
Mouse IgM is purified from ascites or culture
supernatants by an IgM affinity
matrix column.
Ion Exchange
This column can be employed to purify antibodies preceded
by ammonium sulfate
fractionation or followed by affinity chromatography. The ion exchange method is particularly useful for
isolating antibodies that have weak binding affinity to Protein A, or removing traces of DNA contamination.
Additional service: Agarose Bead Conjugation
Our agarose bead
conjugation services offer agarose
bead conjugations to antibody molecules for applications like antibody affinity purification and
immuno-precipitation.
Our customer service representatives are available 24 hours Monday through Friday. You may contact us anytime for assistance. Orders can be placed online, over the phone, by email or fax. Please have your payment information (credit card, purchase order or bank information) ready.
