Development of a highly sensitive chemical method for detecting alpha2-->8-linked oligo/polysialic acid residues in glycoproteins blotted on the membrane

Anal Biochem. 1998 Aug 1;261(2):191-7. doi: 10.1006/abio.1998.2718.

Abstract

A highly sensitive chemical method for detecting alpha2-->8-linked oligo/polysialic acid (oligo/polySia) chains was developed, including (i) periodate oxidation, reduction with sodium borohydride, and subsequent acid hydrolysis, giving rise to C7 analogues and intact C9 compounds from nonreducing terminal and internal sialic acid residues, respectively; (ii) fluorescent labeling of these C7 and C9 compounds with 1,2-diamino-4,5-methylenedioxybenzene (DMB); and (iii) quantitation of theseDMB derivatives on fluorometric high-performance liquid chromatography. As little as 1 ng of internal sialic acid residues of oligo/polySia chains, the existence of which indicates the presence of oligo/polySia structure, was detectable by this method. This fluorometric C7/C9 analysis was successfully applied to glycoproteins blotted onto a slit of polyvinylidene fluoride membranes and suggested the presence of some novel oligoSia-containing glycoproteins in pig embryonic brains.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Brain / embryology
  • Brain Chemistry
  • Chromatography, High Pressure Liquid
  • Fluorescent Dyes
  • Glycoproteins / chemistry*
  • Phenylenediamines
  • Polyvinyls
  • Sensitivity and Specificity
  • Sialic Acids / analysis*
  • Sialic Acids / chemistry
  • Spectrometry, Fluorescence / methods*
  • Swine

Substances

  • Fluorescent Dyes
  • Glycoproteins
  • Phenylenediamines
  • Polyvinyls
  • Sialic Acids
  • polysialic acid
  • polyvinylidene fluoride
  • 1,2-diamino-4,5-methylenedioxybenzene