Specificity analysis of lectins and antibodies using remodeled glycoproteins

Thomas Iskratsch, Andreas Braun, Katharina Paschinger, Iain B. H. Wilson

Research output: Contribution to journalArticlepeer-review

104 Citations (Scopus)
1 Downloads (Pure)

Abstract

Due to their ability to bind specifically to certain carbohydrate sequences, lectins are a frequently used tool in cytology, histology, and glycan analysis but also offer new options for drug targeting and drug delivery systems. For these and other potential applications, it is necessary to be certain as to the carbohydrate structures interacting with the lectin. Therefore, we used glycoproteins remodeled with glycosyltransferases and glycosidases for testing specificities of lectins from Aleuria aurantia (AAL), Erythrina cristagalli (ECL), Griffonia simplicifolia (GSL I-B(4)), Helix pomatia agglutinin (HPA), Lens culinaris (LCA), Lotus tetragonolobus (LTA), peanut (Arachis hypogaeae) (PNA), Ricinus communis (RCA I), Sambucus nigra (SNA), Vicia villosa (VVA), and wheat germ (Triticum vulgaris) (WGA) as well as reactivities of anti-carbohydrate antibodies (anti-bee venom, anti-horseradish peroxidase [anti-HRP], and anti-Lewis(x)). After enzymatic remodeling, the resulting neoglycoforms display defined carbohydrate sequences and can be used, when spotted on nitrocellulose or in enzyme-linked lectinosorbent assays, to identify the sugar moieties bound by the lectins. Transferrin with its two biantennary complex N-glycans was used as scaffold for gaining diverse N-glycosidic structures, whereas fetuin was modified using glycosidases to test the specificities of lectins toward both N- and O-glycans. In addition, alpha(1)-acid glycoprotein and Schistosoma mansoni egg extract were chosen as controls for lectin interactions with fucosylated glycans (Lewis(x) and core alpha1,3-fucose). Our data complement and expand the existing knowledge about the binding specificity of a range of commercially available lectins.

Original languageEnglish
Pages (from-to)133-146
Number of pages14
JournalAnalytical Biochemistry
Volume386
Issue number2
DOIs
Publication statusPublished - 15 Mar 2009

Keywords

  • Antibodies, Monoclonal
  • Antibody Specificity
  • Binding Sites
  • Clinical Laboratory Techniques
  • Fucose
  • Glycoproteins
  • Humans
  • Lectins
  • Plant Lectins
  • Polysaccharides

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