Facile MALDI-MS analysis of neutral glycans in NaOH-doped matrixes: microwave-assisted deglycosylation and one-step purification with diamond nanoparticles.
A streamlined protocol has been developed to accelerate, simplify, and enhance matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) of neutral underivatized glycans released from glycoproteins. It involved microwave-assisted enzymatic digestion and release of glycans, followed by rapid removal of proteins and peptides with carboxylated/oxidized diamond nanoparticles, and finally treating the analytes with NaOH before mixing them with acidic matrix (such as 2,5-dihydroxybenzoic acid) to suppress the formation of both peptide and potassiated oligosaccharide ions in MS analysis. The advantages of this protocol were demonstrated with MALDI-TOF-MS of N-linked glycans released from ovalbumin and ribonuclease B.
Tzeng
Yan-Kai
YK
Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 106, Taiwan.
Chang
Cheng-Chun
CC
Huang
Chien-Ning
CN
Wu
Chih-Che
CC
Han
Chau-Chung
CC
Chang
Huan-Cheng
HC
D018130:Diamond / Q000737:chemistry*; D006023:Glycoproteins / Q000378:metabolism; D006031:Glycosylation; D008872:Microwaves; D053758:Nanoparticles / Q000737:chemistry*; D009844:Oligosaccharides / Q000737:chemistry; D010084:Oxidation-Reduction; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012972:Sodium Hydroxide / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism
D006023:Glycoproteins; D009844:Oligosaccharides; D011134:Polysaccharides; D012972:Sodium Hydroxide; D018130:Diamond; D014357:Trypsin; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase