High-efficiency labeling of sialylated glycoproteins on living cells

Nat Methods. 2009 Mar;6(3):207-9. doi: 10.1038/nmeth.1305. Epub 2009 Feb 22.

Abstract

We describe a simple method for efficiently labeling cell-surface sialic acid-containing glycans on living animal cells. The method uses mild periodate oxidation to generate an aldehyde on sialic acids, followed by aniline-catalyzed oxime ligation with a suitable tag. Aniline catalysis dramatically accelerates oxime ligation, allowing use of low concentrations of aminooxy-biotin at neutral pH to label the majority of cell-surface sialylated glycoproteins while maintaining high cell viability.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aldehydes / chemistry
  • Aniline Compounds / chemistry
  • Animals
  • Biotin / analogs & derivatives
  • Biotin / chemistry
  • Biotinylation / methods*
  • CHO Cells
  • Catalysis
  • Cell Line
  • Cell Survival
  • Cricetinae
  • Cricetulus
  • Endocytosis
  • Flow Cytometry
  • Humans
  • Leukocyte Common Antigens / chemistry
  • Membrane Glycoproteins / chemistry*
  • Microscopy, Fluorescence
  • N-Acetylneuraminic Acid / analogs & derivatives
  • N-Acetylneuraminic Acid / chemistry*
  • Oxidation-Reduction
  • Oximes / chemistry
  • Periodic Acid / chemistry
  • Sialyltransferases / chemistry
  • Staining and Labeling / methods*
  • Streptavidin / chemistry

Substances

  • Aldehydes
  • Aniline Compounds
  • Membrane Glycoproteins
  • Oximes
  • Periodic Acid
  • Biotin
  • Streptavidin
  • metaperiodate
  • Sialyltransferases
  • beta-D-galactoside alpha 2-6-sialyltransferase
  • Leukocyte Common Antigens
  • PTPRC protein, human
  • N-Acetylneuraminic Acid
  • aniline