Antibody containing supernatants are extensively clarified prior to purification to remove all cell debris and reduce the presence of interfering molecules such as DNA and proteases. This enables a very effective one step purification process.
Purification of IgG is carried out by protein A or protein G affinity purification. We also have protocols for purification of IgMs.
Protein A and Protein G are microbial proteins expressed on the cell surface of Staphylococcal and Streptococcal bacteria respectively, and function to bind host immunoglobulin at Fc regions. These binding properties also make them effective tools for antibody purification when used in chromatography systems.
Clarified supernatants are run through columns containing protein A/G immobilized on agarose beads, using a chromatography system. As the supernatant passes through the system, the antibody is bound to Protein A/G. The bound antibody can then be eluted from the column using acidic elution buffer, releasing the final purified product
Once purified the monoclonal antibody is buffer exchanged and gently concentrated to the required concentration before filter sterilising and vialing as required.
We can provide a certificate of analysis of the final product based on your parameters e.g. ELISA, SDS-PAGE, Western Blot, endotoxin level, sterility etc.