16
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Tandem mass spectrometry Mass spectrometry for analyses of Post-translational modification
1.Tandem in space (空間性串聯) 2.Tandem in time (時間性串聯)
LID
TOF-TOF
Ion Trap
Ion Trap Radio frequency field is scanned multiple collision-induced dissociation
1
Protein Phosphorylation
Parent ion scan (Precursor ion scan)
2
Analysis of phosphorylated positions
3
Protein glycosylation
4
5
Glycoproteins
glycoform
6
N-linked oligosaccharide core Enzyme release
Oligomannose-type N-linked oligosaccharides
O-linked oligosaccharide core
Hybrid and complex native N-linked oligosaccharides
O-linked oligosaccharide core
Hydrazine release
7
Functions of glycoproteins
C3 convertase
Methods for analyses of glycan
bind to C8 and C9
Analyses of glycan by liquid chromatography
8
O-linked oligosaccharide core
N-linked oligosaccharide core
N-glycosidase F
2-AB labeling
HPLC 1. Normal phase HPLC (NP) Glucose uint (dextran)
2. Reverse phase HPLC (RP) Arabinose unit 3. Weak Anionic Exchanger (WAX)
1. Normal phase HPLC (NP) Glucose uint (dextran)
2. Reverse phase HPLC (RP) Arabinose unit 3. Weak Anionic Exchanger (WAX)
9
1. Normal phase HPLC (NP) Glucose uint (dextran)
2. Reverse phase HPLC (RP) Arabinose unit 3. Weak Anionic Exchanger (WAX)
Enzyme array
2D electrophoresis and MALDI-TOF MS
10
Glycoform ?
MALDI-TOF and glycoform MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight)
N-glycosidase F
11
Ion Trap
Tandem mass spectrometry (MSn)
Ion Trap
12
Chemical modification
13
14
1.Tandem in space (空間性串聯) 2.Tandem in time (時間性串聯)
LID
TOF-TOF
Ion Trap
Ion Trap Radio frequency field is scanned multiple collision-induced dissociation
1
Protein Phosphorylation
Parent ion scan (Precursor ion scan)
2
Analysis of phosphorylated positions
3
Protein glycosylation
4
5
Glycoproteins
glycoform
6
N-linked oligosaccharide core Enzyme release
Oligomannose-type N-linked oligosaccharides
O-linked oligosaccharide core
Hybrid and complex native N-linked oligosaccharides
O-linked oligosaccharide core
Hydrazine release
7
Functions of glycoproteins
C3 convertase
Methods for analyses of glycan
bind to C8 and C9
Analyses of glycan by liquid chromatography
8
O-linked oligosaccharide core
N-linked oligosaccharide core
N-glycosidase F
2-AB labeling
HPLC 1. Normal phase HPLC (NP) Glucose uint (dextran)
2. Reverse phase HPLC (RP) Arabinose unit 3. Weak Anionic Exchanger (WAX)
1. Normal phase HPLC (NP) Glucose uint (dextran)
2. Reverse phase HPLC (RP) Arabinose unit 3. Weak Anionic Exchanger (WAX)
9
1. Normal phase HPLC (NP) Glucose uint (dextran)
2. Reverse phase HPLC (RP) Arabinose unit 3. Weak Anionic Exchanger (WAX)
Enzyme array
2D electrophoresis and MALDI-TOF MS
10
Glycoform ?
MALDI-TOF and glycoform MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight)
N-glycosidase F
11
Ion Trap
Tandem mass spectrometry (MSn)
Ion Trap
12
Chemical modification
13
14
