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. 2022:2442:55-74.
doi: 10.1007/978-1-0716-2055-7_3.

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

Anu Paul #  1 , Shang-Chuen Wu #  1 , Kashyap R Patel  1 , Alex D Ho  1 , Jerry William Lynn Allen  1 , Hans Verkerke  1 , Connie M Arthur  1   2 , Sean R Stowell  3   4
Affiliations

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

Anu Paul et al. Methods Mol Biol. 2022.

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.

Keywords: Coprinopsis cinerea galectin-2 (CGL2); Divinyl sulfone-coupled lactosyl-Sepharose; Glutathione-Sepharose affinity purification of GST-tagged human galectin-7; Lactosyl-Sepharose affinity chromatography; Nickel NTA-affinity purification of histidine-tagged human galectin-7; Recombinant galectin purification.

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