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Production of Recombinant Proteins from Pichia pastoris: Interfacing Fermentation and Immobilized Metal Ion Affinity Chromatography

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Laboratory Protocols in Fungal Biology

Part of the book series: Fungal Biology ((FUNGBIO))

Abstract

The methods describe a Pichia pastoris fermentation system for ­generation and purification of recombinant proteins. The proteins are secreted with hexahistidine tags and purified from feedstock by immobilized metal ion affinity chromatography (IMAC) using either radial flow or expanded bed adsorption. IMAC allows for an initial fast capture and isolation step that omits the need for filtration or centrifugation as primary procedures. The methods are applicable to production of recombinant protein in the laboratory and can be adapted to good manufacturing practice (GMP) compliant processes.

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Notes

  1. 1.

     e.g., http://www2.dupont.com/DAHS_EMEA/en_GB/products/disinfectants/virkon_s/index.htmL.

  2. 2.

    e.g., http://www.newpig.com.

  3. 3.

     http://www.nalgenelabware.com/techdata/technical/FiltrationIndex.asp.

  4. 4.

    http://www.millipore.com/catalogue/item/xx42lss11.

  5. 5.

    http://www.pall.com/laboratory_53238.asp.

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Acknowledgements

This work was supported by Cancer Research UK; Department of Health (ECMC, Experimental Cancer Medicine Network Centre); Engineering and Physical Sciences Research Council (EPSRC); The Breast Cancer Campaign; and UCL Cancer Institute Research Trust.

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Authors and Affiliations

  1. Department of Oncology, University College London Cancer Institute, Paul O’Gorman Building, 72 Huntley Street, London, WC1E 6BT, UK

    Berend Tolner, Gaurav Bhavsar, Bride Foster, Kim Vigor & Kerry Chester

Authors
  1. Berend Tolner
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  2. Gaurav Bhavsar
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  3. Bride Foster
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  4. Kim Vigor
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  5. Kerry Chester
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Corresponding author

Correspondence to Berend Tolner .

Editor information

Editors and Affiliations

  1. , Department of Biochemistry, National University of Ireland Galway, University Road, Galway, N/A, Ireland

    Vijai Kumar Gupta

  2. , Department of Biochemistry, National University of Ireland Galway, University Road, Galway, N/A, Ireland

    Maria G. Tuohy

  3. , Department of Biochemistry, National University of Ireland Galway, University Road, Galway, N/A, Ireland

    Manimaran Ayyachamy

  4. , Manufacturing Sciences and Technology, Pfizer Ireland Pharmaceuticals, The Pfizer Biotech Campus at Grange Castle, Clondalkin, N/A, Dublin, Ireland

    Kevin M. Turner

  5. , School of Natural Sciences, National University of Ireland Galway, University Road, Galway, N/A, Ireland

    Anthonia O’Donovan

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Tolner, B., Bhavsar, G., Foster, B., Vigor, K., Chester, K. (2013). Production of Recombinant Proteins from Pichia pastoris: Interfacing Fermentation and Immobilized Metal Ion Affinity Chromatography. In: Gupta, V., Tuohy, M., Ayyachamy, M., Turner, K., O’Donovan, A. (eds) Laboratory Protocols in Fungal Biology. Fungal Biology. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-2356-0_37

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