Integrated isolation of antibody fragments from microbial cell culture fluids using supermacroporous cryogels

Journal of Chromatography. a
Maria B DainiakBo Mattiasson

Abstract

The present paper describes a chromatographic capture/purification step for the recovery of proteins directly from undiluted and unclarified cell culture broths using supermacroporous dimethylacrylamide (DMAA) cryogel. The interconnected character and the size (10-100 microm) of the pores of the adsorbent make it possible to process whole cell fermentation broths without blocking the column. Cu2+-iminodiacetic acid (IDA) DMAA cryogel has been used for the isolation and purification of excreted (His)6-tagged single chain (sc) Fv antibody fragments, (His)6-scFv, from E. coli cell culture. Bound protein was recovered with 0.2 M imidazole or with 20 mM EDTA and was practically cell-free. Chromatographic capture using Cu2+-IDA cryogel column was performed at flow rates of 300 and 600 cm/h, respectively and resulted in 84-96% recovery of (His)6-scFv fragments with a purification factor of 13-15. The DMAA cryogel adsorbent is mechanically stable, can withstand harsh cleaning-in-place procedure and is relatively inexpensive. Chromatographic isolation of proteins using cryogels allows efficient removal of cells and can be operated at a flow rate as high as 600 cm/h. This novel technique has proven to be a scalable process, does not requ...Continue Reading

References

Jun 5, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Fatima M PlievaBo Mattiasson

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Citations

Sep 17, 2010·Journal of Biomaterials Science. Polymer Edition·Deepti SinghAshok Kumar
Oct 13, 2007·Journal of Biomaterials Science. Polymer Edition·Nimet BölgenErhan Pişkin
Jul 10, 2010·International Journal of Biological Sciences·Deepti SinghAshok Kumar
Jan 19, 2006·Proceedings of the National Academy of Sciences of the United States of America·Maria B DainiakBo Mattiasson
Aug 13, 2013·BioMed Research International·Archana SharmaAshok Kumar
Jan 13, 2016·Artificial Cells, Nanomedicine, and Biotechnology·Hüseyin AlkanMehmet Odabaşı
Oct 9, 2012·Enzyme and Microbial Technology·Akshay SrivastavaAshok Kumar
Jul 24, 2012·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Poondi Rajesh GavaraMarcelo Fernandez-Lahore
Oct 7, 2008·Trends in Microbiology·Fatima M PlievaBo Mattiasson
Dec 1, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Duncan LowNarahari S Pujar
May 1, 2009·Biotechnology and Bioengineering·Jagan M Billakanti, Conan J Fee
Jun 20, 2008·Journal of Biomedical Materials Research. Part a·Anuj TripathiAshok Kumar
Aug 12, 2008·Journal of Biomedical Materials Research. Part a·Nimet BölgenErhan Pişkin
Oct 18, 2011·Journal of Tissue Engineering and Regenerative Medicine·N BölgenE Pişkin
Jun 22, 2011·Biotechnology Progress·Noor Shad BibiMarcelo Fernández-Lahore
May 8, 2013·Biotechnology Journal·Telma BarrosoAna Aguiar-Ricardo
Oct 12, 2015·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Müge AndaçAdil Denizli
Aug 2, 2005·Protein Expression and Purification·Vibha Bansal, Pradip K Roychoudhury
Jun 12, 2014·Journal of Chromatography. a·Gizem Ertürk, Bo Mattiasson
Apr 11, 2015·Journal of Chromatography. a·Willer Ferreira da Silva JúniorBruna Mara Aparecida de Carvalho
Jul 1, 2011·Journal of Hazardous Materials·Irina N SavinaAndrew B Cundy
Aug 14, 2019·Journal of Biomedical Materials Research. Part a·Mehdi RazaviAvnesh S Thakor
Jan 12, 2013·Artificial Cells, Nanomedicine, and Biotechnology·Şeyda Ceylan, Mehmet Odabaşı

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