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Magnetic Precipitation: A New Platform for Protein Purification
Publication . Dos Santos, Raquel; Iria, Inês; Manuel, Ana M.; Leandro, Ana P.; Madeira, Catarina A. C.; Goncalves, João; Carvalho, Ana Luísa; Roque, Ana Cecília; DQ - Departamento de Química; UCIBIO - Applied Molecular Biosciences Unit; Wiley
One of the trends in downstream processing comprises the use of “anything-but-chromatography” methods to overcome the current downfalls of standard packed-bed chromatography. Precipitation and magnetic separation are two techniques already proven to accomplish protein purification from complex media, yet never used in synergy. With the aim to capture antibodies directly from crude extracts, a new approach combining precipitation and magnetic separation is developed and named as affinity magnetic precipitation. A precipitation screening, based on the Hofmeister series, and a commercial precipitation kit are tested with affinity magnetic particles to assess the best condition for antibody capture from human serum plasma and clarified cell supernatant. The best conditions are obtained when using PEG3350 as precipitant at 4 °C for 1 h, reaching 80% purity and 50% recovery of polyclonal antibodies from plasma, and 99% purity with 97% recovery yield of anti-TNFα mAb from cell supernatants. These results show that the synergetic use of precipitation and magnetic separation can represent an alternative for the efficient capture of antibodies.
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Fundação para a Ciência e a Tecnologia
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3599-PPCDT
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RECI/BBB-BEP/0124/2012