二氧化钛结合超滤膜富集和分离肿瘤患者唾液中的磷酸化肽和唾液酸化糖肽

doi:10.7503/cjcu20140659

摘要/Abstract

摘要：

利用TiO₂富集、超滤膜滤过实现了选择性富集和分离磷酸化肽和含唾液酸的N-链接糖肽. 首先选取截留分子量为1×10⁴的超滤膜分离糖肽和磷酸化肽, 然后利用酪蛋白和牛血清白蛋白的酶解物验证所建立的方法, 该方法的最低检出限是0.16 pmol. 将上述方法应用于肺癌患者的唾液检测, 成功检测并鉴定到8个磷酸化肽和5个糖肽. 本方法可有效提高磷酸化肽和糖肽检测的选择性和灵敏度, 为疾病生物标志物的检测提供了新方法.

关键词: 二氧化钛, 富集, 超滤膜, 磷酸化肽, 糖肽

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

中图分类号:

O657.7

TrendMD:

李娟, 姜武辉, 徐晓颖. 二氧化钛结合超滤膜富集和分离肿瘤患者唾液中的磷酸化肽和唾液酸化糖肽. 高等学校化学学报, 2014, 35(10): 2073.

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^†. Chem. J. Chinese Universities, 2014, 35(10): 2073.

图/表 4

参考文献 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