Challenging the limits of detection of sialylated Thomsen-Friedenreich antigens by in-gel deglycosylation and nano-LC-MALDI-TOF-MS

• Type: Article in International Scientific Journal
• Date: 2013

Authors

Andreia Almeida (Author) Other; Jose A Ferreira (Author) Other; Filipe Teixeira (Author) Other; Catarina Gomes (Author) Other; Natalia N D S Cordeiro (Author) FCUP; Hugo Osorio (Author) Other; Lucio Lara Santos (Author) ICBAS; Celso A Reis (Author) ICBAS; Rui Vitorino (Author) Other; Francisco Amado (Author) Other

Journal

Title: Electrophoresis

Vol. 34

Pages: 2337-2341

ISSN: 0173-0835

Publisher: Wiley-Blackwell

Indexing

ISI Web of Knowledge - 11 Citations

Scopus - 12 Citations

Scientific classification

FOS: Natural sciences > Chemical sciences

Other information

Authenticus ID: P-006-K24

DOI: 10.1002/elps.201300148

Abstract (EN): The identification of sialylated Thomsen-Friedenreich antigens in proteins poses much interest in the context of cancer research. MALDI-TOF-MS is a powerful technique for this purpose; still it shows considerable low sensitivity for sialylated molecules due to in-source and metastable decomposition. Herein, we report a target-driven strategy to identify these antigens in minute amounts of glycoproteins isolated in polyacrylamide gels. The glycans were recovered from gel spots by reductive -elimination, permethylated and analyzed by nano-LC-MALDI-TOF-MS. A computational algorithm was developed to filter spectral noise and enhance/isolate the signals of interest. Sialylated antigens were identified in minute amounts of fetuin (0.1 g) and plasminogen (1.0 g) by this approach.MS assignments were further validated by enzymatic methods. This methodology allowed a fivefold decrease in the current LOD of fetuin sialylated O-glycans by MALDI-TOF-MS.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 5