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Purification of Recombinant Galectins from Different Species Using Distinct Affinity Chromatography Methods

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Galectins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2442))

Abstract

Galectins are lectins having the capacity to recognize β-galactose-containing glycan structures and are widely distributed among various taxa. However, the exact physiological and biochemical functions mediated by galectins that necessitate their wide occurrence among diverse species have not yet been delineated in a precise manner. Purification of recombinant galectins in active form is a fundamental requirement to elucidate their biological function. In this chapter, we are describing methods to recombinantly express and purify galectins using three different methods of affinity purification, i.e., lactosyl-Sepharose chromatography for fungal galectin Coprinopsis cinerea galectin 2 (CGL2), nickel-chromatography for histidine-tagged human galectin-7, and glutathione-Sepharose chromatography for Glutathione S-transferase-tagged (GST-tagged) human galectin-7. Step-by-step instructions are provided for obtaining the above-mentioned recombinant galectins that retain carbohydrate-binding activity and are suitable for conducting biochemical experiments.

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Acknowledgments

This work was supported in part by National Institutes of Health Grants R01 HL154034 to CMA and U01 CA242109 to SRS.

Conflict of Interest

No conflict of interest declared.

Author information

Author notes

  1. Anu Paul and Shang-Chuen Wu contributed equally with all other contributors.

Authors and Affiliations

  1. Joint Program in Transfusion Medicine, Department of Pathology, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Anu Paul, Shang-Chuen Wu, Kashyap R. Patel, Alex D. Ho, Jerry William Lynn Allen, Hans Verkerke, Connie M. Arthur & Sean R. Stowell

  2. Harvard Glycomics Center, Harvard Medical School, Boston, MA, USA

    Connie M. Arthur & Sean R. Stowell

Authors
  1. Anu Paul
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  2. Shang-Chuen Wu
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  3. Kashyap R. Patel
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  4. Alex D. Ho
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  5. Jerry William Lynn Allen
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  6. Hans Verkerke
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  7. Connie M. Arthur
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  8. Sean R. Stowell
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Corresponding author

Correspondence to Sean R. Stowell .

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Editors and Affiliations

  1. Joint Program in Transfusion Medicine Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Sean R. Stowell

  2. Joint Program in Transfusion Medicine Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Connie M. Arthur

  3. Department of Surgery Beth Israel Deaconess Medical Center National Center for Functional Glycomics, Harvard Medical School, Boston, MA, USA

    Richard D. Cummings

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Paul, A. et al. (2022). Purification of Recombinant Galectins from Different Species Using Distinct Affinity Chromatography Methods. In: Stowell, S.R., Arthur, C.M., Cummings, R.D. (eds) Galectins. Methods in Molecular Biology, vol 2442. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2055-7_3

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  • DOI: https://doi.org/10.1007/978-1-0716-2055-7_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2054-0

  • Online ISBN: 978-1-0716-2055-7

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