Boronic acid lectin affinity chromatography (BLAC). 2. Affinity micropartitioning-mediated comparative glycosylation profiling

Alex Monzo, Marcell Olajos, Lorenzo De Benedictis, Zuly Rivera, Guenther K. Bonn, A. Guttman

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

As a continuation of our work on boronic acid lectin affinity chromatography (BLAC), in this paper we introduce an automated affinity micropartitioning approach using combined boronic acid and concanavalin A (BLAC/Con A) resin-filled micropipette tips to isolate and enrich human serum glycoproteins. The N-linked oligosaccharides of the partitioned glycoproteins were removed by PNGase F enzyme digestion, followed by 8-aminopyrene-1,3, 6-trisulfonic acid labeling. Capillary gel electrophoresis with blue LED-induced fluorescence detection was applied in a multiplexed format for comparative glycan profiling. The efficiency of BLAC affinity micropartitioning was compared with that of the individual lectin and pseudolectin affinity enrichment. Finally, we report on our findings in glycosylation differences in human serum samples from healthy and prostate cancer patients by applying BLAC/Con A micropipette tip-based enrichment and comparative multicapillary gel electrophoresis analysis of the released and labeled glycans.

Original languageEnglish
Pages (from-to)195-201
Number of pages7
JournalAnalytical and Bioanalytical Chemistry
Volume392
Issue number1-2
DOIs
Publication statusPublished - Sep 2008

Fingerprint

Boronic Acids
Glycosylation
Affinity chromatography
Affinity Chromatography
Lectins
Electrophoresis
Polysaccharides
Glycoproteins
Gels
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
Capillary Electrophoresis
Concanavalin A
Serum
Oligosaccharides
Labeling
Light emitting diodes
Digestion
Prostatic Neoplasms
Resins
Fluorescence

Keywords

  • Affinity chromatography
  • Capillary gel electrophoresis
  • Glycan profiling

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Cite this

Boronic acid lectin affinity chromatography (BLAC). 2. Affinity micropartitioning-mediated comparative glycosylation profiling. / Monzo, Alex; Olajos, Marcell; De Benedictis, Lorenzo; Rivera, Zuly; Bonn, Guenther K.; Guttman, A.

In: Analytical and Bioanalytical Chemistry, Vol. 392, No. 1-2, 09.2008, p. 195-201.

Research output: Contribution to journalArticle

Monzo, Alex ; Olajos, Marcell ; De Benedictis, Lorenzo ; Rivera, Zuly ; Bonn, Guenther K. ; Guttman, A. / Boronic acid lectin affinity chromatography (BLAC). 2. Affinity micropartitioning-mediated comparative glycosylation profiling. In: Analytical and Bioanalytical Chemistry. 2008 ; Vol. 392, No. 1-2. pp. 195-201.
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