Generation and Use of Recombinant Galectins

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

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Galectins are soluble carbohydrate binding proteins that can bind β-galactose-containing glycoconjugates by means of a conserved carbohydrate recognition domain (CRD). In mammalian systems, galectins have been shown to mediate very important roles in innate and adaptive immunity as well as facilitating host-pathogen relationships. Many of these studies have relied on purified recombinant galectins to uncover key features of galectin biology. A major limitation to this approach is that certain recombinant galectins purified using standard protocols are easily susceptible to loss of glycan-binding activity. As a result, biochemical studies that employ recombinant galectins can be misleading if the overall activity of a galectin remains unknown in a given assay condition. This article examines fundamental considerations when purifying galectins by lactosyl-sepharose and nickel-NTA affinity chromatography using human galectin-4N and -7 as examples, respectively. As other approaches are also commonly applied to galectin purification, we also discuss alternative strategies to galectin purification, using human galectin-1 and -9 as examples.

Original languageEnglish
Article numbere63
JournalCurrent Protocols
Issue number3
StatePublished - Mar 2021
Externally publishedYes


  • galectin
  • galectin stability
  • galectin-1 alkylation using iodoacetamide
  • galectin-4
  • galectin-7
  • galectin-9 purification using Tris buffer
  • lactosyl-sepharose chromatography
  • stable and active galectin-9


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