What is Monoclonal Antibody Purification?
Monoclonal antibody purification involves collecting the media from the cultured hybridomas, centrifuging and filtering the media, and then using chromatographic or physiochemical fractionation methods to isolate the antibodies. Physiochemical fractionation involves using differential precipitation, size-exclusion or solid-phase binding of proteins based on size, charge or other chemical characteristics. These techniques include Size Exclusion Chromotography (SEC), ammonium sulfate precipitation and Ion Exchange Chromotography. Antibodies can also be purified based on class-specific affinity which involves using immobilized agents, such as proteins or lectins, that have an affinity for the isotype of the antibody of interest. Antigen-affinity chromatography can also be used to purify antibodies, but this method is not commonly used for monoclonal antibodies since the target antibody is generally the only immunoglobulin in the sample.