Novel Method Coupled TiO2 with Ultrafiltration Membrane to Enrich and Separate Phosphopeptides and Sialic Acid-containing Glycopeptides from Saliva of Cancer Patient†

doi:10.7503/cjcu20140659

Abstract

Abstract:

A novel method coupled TiO₂ with ultrafiltration membrane was established to enrich and separate phosphopeptides and N-linked glycopeptides. Firstly, the 10K cutoff ultrafiltration membrane was chosen for separating phosphopeptides and glycopeptides. Then, TiO₂ enrichment and ultrafiltration membrane separation of PTM-peptides were tested with tryptic digests of α-casein and bovine fetuin. The limit of detection of the developed method was measured as 0.16 pmol with standard phosphopeptide. At last, the established method was applied to enrich and separate phosphopeptides and glycopeptides from saliva of lung cancer. A total of 8 phosphopeptides and 5 glycopeptides could be successfully identified with nano ultra-high pressure liquid chromatography-quadrupole-time of flight MS(Nano UPLC Q-TOF). This method significantly improved the sensitivity and efficiency and could be contributed to novel disease biomarker discovery.

Key words: TiO2, Enrichment, Ultrafiltration Membrane, Phosphopeptide, Glycopeptide

CLC Number:

O657.7

TrendMD:

LI Juan, JIANG Wuhui, XU Xiaoying. Novel Method Coupled TiO₂ with Ultrafiltration Membrane to Enrich and Separate Phosphopeptides and Sialic Acid-containing Glycopeptides from Saliva of Cancer Patient^†[J]. Chem. J. Chinese Universities, 2014, 35(10): 2073.

Figures/Tables 4

References 17

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No.	Protein name	Sequence^*	m/z(charge state)	Score
1	Immunoglobulin heavy chain	GLEWIGEIHPSGSTGYNPSLEpSR	856.0339(3+)	27
2	Immunoglobulin kappa chain variable region	TYPIpTFGQGTR	660.8475(2+)	34
3	Aminopeptidase puromycin sensitive, isoform CRA_a	LIKLpSVEGFAVDK	749.788(2+)	29
4	Cystatin S acidic isoform	SSpSKEENR	508.6975(2+)	35
5	Chain 1	NS(ph)DIAFLIDGSGpSIIPHDFRR	830.7573(3+)	42
6	D40	QQLFAApTNR	564.8474(2+)	26
7	Nck-associated protein 1 isoform 2	HHIGDRpSLSLCNMFLDEMAK	824.0644(3+)	56
8	HSPC052	VTQGKEEpTR	564.348(2+)	30
9	Complement factor H	ISEENETTCYMGK	1561.6276(2+)	42
10	Alpha-1 protease inhibitor	YIGNATAIFFIPDEGK	1755.872(2+)	36
11	Haptoglobin	VVLHPNYSKVDIGLIK	898.4424(2+)	60
12	Ig mu chain C region	GITFQQNASSMCVPDQDTAIR	2355.0471(2+)	56
13	Histidine-proline-rich glycoprotein	VIDFNCTTSSVSSAIANTK	2014.9517(2+)	86

No.	Protein name	Sequence^*	m/z(charge state)	Score
1	Immunoglobulin heavy chain	GLEWIGEIHPSGSTGYNPSLEpSR	856.0339(3+)	27
2	Immunoglobulin kappa chain variable region	TYPIpTFGQGTR	660.8475(2+)	34
3	Aminopeptidase puromycin sensitive, isoform CRA_a	LIKLpSVEGFAVDK	749.788(2+)	29
4	Cystatin S acidic isoform	SSpSKEENR	508.6975(2+)	35
5	Chain 1	NS(ph)DIAFLIDGSGpSIIPHDFRR	830.7573(3+)	42
6	D40	QQLFAApTNR	564.8474(2+)	26
7	Nck-associated protein 1 isoform 2	HHIGDRpSLSLCNMFLDEMAK	824.0644(3+)	56
8	HSPC052	VTQGKEEpTR	564.348(2+)	30
9	Complement factor H	ISEENETTCYMGK	1561.6276(2+)	42
10	Alpha-1 protease inhibitor	YIGNATAIFFIPDEGK	1755.872(2+)	36
11	Haptoglobin	VVLHPNYSKVDIGLIK	898.4424(2+)	60
12	Ig mu chain C region	GITFQQNASSMCVPDQDTAIR	2355.0471(2+)	56
13	Histidine-proline-rich glycoprotein	VIDFNCTTSSVSSAIANTK	2014.9517(2+)	86

Novel Method Coupled TiO₂ with Ultrafiltration Membrane to Enrich and Separate Phosphopeptides and Sialic Acid-containing Glycopeptides from Saliva of Cancer Patient^†

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