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

被引：9

作者：

Almeida, Andreia ^{[1
]}

Ferreira, Jose A. ^{[1
,2
]}

Teixeira, Filipe ^{[3
]}

Gomes, Catarina ^{[4
]}

Cordeiro, M. Natalia D. S. ^{[3
]}

Osorio, Hugo ^{[4
,5
]}

Santos, Lucio Lara ^{[2
,6
,7
]}

Reis, Celso A. ^{[4
,5
,8
]}

Vitorino, Rui ^{[1
]}

Amado, Francisco ^{[1
,9
]}

机构：

[1] Univ Aveiro, Dept Chem, Mass Spectrometry Ctr, QOPNA, P-3810193 Aveiro, Portugal

[2] Portuguese Inst Oncol, Expt Pathol & Therapeut Grp, Oporto, Portugal

[3] Univ Porto, Dept Chem & Biochem, Fac Sci, REQUIMTE, P-4100 Oporto, Portugal

[4] Univ Porto IPATIMUP, Inst Mol Pathol & Immunol, Oporto, Portugal

[5] Univ Porto, Fac Med, P-4100 Oporto, Portugal

[6] Univ Fernando Pessoa, Hlth Sch, Oporto, Portugal

[7] Portuguese Inst Oncol, Dept Surg Oncol, Oporto, Portugal

[8] Univ Porto, Inst Biomed Sci Abel Salazar, P-4100 Oporto, Portugal

[9] Univ Aveiro ESSUA, Sch Hlth Sci, Aveiro, Portugal

来源：

ELECTROPHORESIS | 2013年 / 34卷 / 16期

关键词：

Nano-LC-MALDI-TOF-MS; Permethylation; Sialic acids; Simple mucin-type antigens; Thomsen-Friedenreich antigens; MASS-SPECTROMETRY; O-GLYCANS; GLYCOPROTEINS; MATRIX; PERMETHYLATION; GLYCOSYLATION; RECOVERY; CELLS; SERUM;

D O I：

10.1002/elps.201300148

中图分类号：

Q5 [生物化学];

学科分类号：

070307 [化学生物学];

摘要：

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.

引用

页码：2337 / 2341

页数：5

共 24 条

[1]

Artemenko Natalia V, 2012, Methods Mol Biol, V899, P325, DOI 10.1007/978-1-61779-921-1_21

[2]

Pathways of O-glycan biosynthesis in cancer cells [J].