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Ready-to-use or activated affinity resins and protein A media are available for affinity purification of affinity-tagged proteins, monoclonal antibodies, and other biomolecules.
Purification of a Monoclonal Antibody Using a Combination of UNOsphere Rapid S and UNOsphere Q Media Ion Exchange Chromatography After Protein A Capture
Introduction Antibody-based drugs represent a growing segment of the pharmaceutical industry and one of the most promising classes of therapeutic drugs. The demand for protein-based drugs is expected to grow steadily for the foreseeable future. However, production of monoclonal antibody (mAb)–based drugs remains very costly, and solutions to lower production costs are needed. Downstream purification steps are becoming a target for cost reduction. The use of ion exchange chromatography to directly capture MAbs from cell culture streams or as a polishing step after an affinity separation may be a way to reduce costs.
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Shrink-swell test on Profinity IMAC resin. Common reagents used in His-tagged protein purifications were run on a 1.1 x 30 cm Amicon column packed with Profinity IMAC resin to a bed height of 20 cm. System pressure and column bed height compression were recorded for each flow rate. Flow rates were increased stepwise to 200 cm/hr and held for 2 min at each step. All tests were performed on a BioLogic DuoFlow Maximizer system. Yellow horizontal line indicates 43 psi (3 bar), the maximum recommended operating pressure.
* 10% breakthrough capacity determined with 1.0 mg/ml polyclonal human IgG in 1.1 x 10 cm column.**No significant change in chromatographic performance
* Compatibility determined using Profinity eXact control lysate; some reagents, like ammonium sulfate, are protein-dependent. ** Chloride ions trigger cleavage of target proteins.
* In order of increasing retention by support.
Bio-Rad offers several protein A media for different mAb purification needs. UNOsphere SUPrA™ Resin is designed for process-scale purification workflows. Affi-Gel® and Affi-Prep® Protein A Resins yield highly purified IgG.
Profinity eXact Affinity Purification Resin consists of a highly engineered subtilisin protease conjugated to an agarose-based matrix.
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