微量复杂生物样品糖蛋白N-糖链电喷雾电离质谱分析前处理方法的建立
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摘要
糖生物学是21世纪生命科学研究的重要组成部分。要深入研究糖链在生命活动中的功能和调控机制,首要任务是获得并分析生物糖组。电喷雾电离质谱(ESI-MS)是目前分析微量糖组的强大工具,但对样品的前处理要求高。本文针对目前存在的问题建立了微量复杂生物样品糖蛋白N-糖链电喷雾电离质谱分析前处理的方法。
本研究先以牛胰核糖核酸酶B和鸡卵清白蛋白二种常见的糖蛋白为研究材料,对几种N-糖链解离、富集纯化方法进行了考察,得到以下结论：
1.N-糖链解离方法：选用十二烷基磺酸钠(SDS)和碘乙酰胺(DTT)作蛋白质的变性剂,100℃反应10 min；加入磷酸钠缓冲液和10%乙基苯基聚乙二醇(NP40),然后加入N-糖苷酶F (PNGase F),37℃反应24 h。这种方法解离效果好,效率最高,操作简单。
2.N-糖链富集纯化方法：我们将富集纯化方法联合使用,对解离样品进行质谱分析前处理,结果表明,除了微晶纤维素和阳离子交换树脂混合装柱法外,其余4种方法(微晶纤维素柱结合阳离子交换树脂,C18柱结合活性炭柱,C18柱结合阳离子交换树脂和活性炭柱结合微晶纤维素柱)均可以达到满意的富集纯化效果。
我们将初步建立的方法用胎牛血清,健康小鼠血清,健康人血清,小鼠HAbI8 IgG以及HAbI8 IgG(腹水法)这5种生物样品进一步验证,并针对微量样品进行了改进：
3.PVDF膜辅助N-糖链解离方法：我们利用PVDF膜能够结合蛋白质的特性,在释放糖链前对复杂生物样品进行粗纯化,再按照上述糖链解离步骤,直接在膜上完成糖蛋白变性和糖链解离过程。实验证明,这种方法能同时提高微量样品糖链纯化效果。
4.改进的N-糖链富集纯化方法：上述4种富集纯化方法中,只有活性炭柱联合微晶纤维素柱纯化生物来源N-糖链解离样品的效果最好。在用活性炭柱进行纯化时,为了避免糖链洗脱不完全,我们用含0.1%三氟乙酸的50%的乙腈溶液为洗脱剂；微晶纤维素柱的平衡时间与填料多少成正比,平衡效果好坏直接影响纯化效果,而微量样品只需要少量的微晶纤维素,因此我们采用1000μL移液器标配枪头作一次性层析柱,可以大大减少干扰物,提高纯化效率。
综上所述,本研究在电喷雾质谱检测技术的基础上,建立了一个更适合于微量生物来源样品糖蛋白N-糖链解离,富集纯化的新方法——采用PVDF膜辅助N-糖链解离方法以及活性炭柱联合微晶纤维素柱的富集纯化方法。这种方法快速,稳定,通用性更强,能够通量化应用于ESI-MS检测前μg级别生物样品糖蛋白N-糖链的制备,有利于糖组的后续研究。
Glycobiology is an important part of life science research in 21st century. To have in-depth study of the functions and regulatory mechanism of glycans in life activities, the primary task is to obtain and analyze glycome. Electrospray Ionization Mass Spectrometry (ESI-MS) is a powerful tool to analyze micro-scale glycomes, but the pretreatment of sample is necessary. In this paper, we established a pretreatment procedure for N-linked glycans released from glycoproteins in micro-scale bio-samples before ESI-MS analysis in view of the exsting problems.
This study firstly used two common glycoproteins, bovine ribonuclease B and chicken albumin, as research materials, and had an initial investigation on several releasing, purifying and enriching N-linked glycans methods and obtained the following conclusions:
1. N-linked glycans releasing:we chose sodium dodecyl sulfate (SDS) and iodine acetamide (DTT) as protein denaturing agents. Glycoproteins were dissolved in the buffer and denatured at 100℃for 10 min. Then added sodium pyrophosphate buffer,10% ethyl-phenyl-polyethyleneglycol (NP40), PNGase F,37℃24 h. This approach is effective, efficient and easy to operate.
2. N-linked glycans purification and enrichment:the enzymatic samples were pretreated separately by five enrichment and purification methods used in conjunction before mass spectrometry analysis. The results showed, in addition to the method of using microcrystalline cellulose and cation exchange resin in tandem, the other four kinds (combination of microcrystalline cellulose and cation exchange resin column, combination of C18 column and graphite carbon column, combination of C18 column and cation exchange resin and combination of graphite carbon column and microcrystalline cellulose column) were able to achieve a satisfactory effect of purification and enrichment.
Then, we used five bio-samples, fetus bovine serum, control mouse serum, control human serum, mouse HAbI8 IgG and HAbI8 IgG (ascites method) for further verification. Moreover, we improved this method focusing on micro-scale samples:
3. PVDF membrane assisted N-linked glycans releasing:the PVDF membrane has the characteristic of absorbing proteins. We used it to roughly purify the complex bio-samples prior to PNGase F digestion, and then in accordance with the above steps completed glycoprotein denaturation and enzymatic digestion directly on the membrane. The results showed that this method can also improve the purification effect of glycans from micro-scale samples.
4. Improved procedure of purifying and enriching.N-linked glycans:In the above mentioned four kinds of enrichment and purification methods, only the combination of graphite carbon column and microcrystalline cellulose column had the best purifying and enriching effect. We used 0.1% trifluoroacetic acid containing 50% acetonitrile solution as eluting agent of graphite carbon column to avoid eluting free glycans incompelely; the pretreatment time of microcrystalline cellulose powder was related with quantities, and the effect of pretreatment would have a direct impact on glycans analysis. The micro-scale samples only required a small amount of powder, so we made use of the 1000μL standard pipettes as the disposable chromatography columns and greatly reduced impurities, improved the efficiency of purification.
This study, based on ESI-MS technology, established a new procedure which was more suitable for purifying and enriching N-linked glycans from micro-scale bio-samples glycoproteins--PVDF membrane assisted N-linked glycans releasing, purifying and enriching free glycans by graphite carbon column and microcrystalline cellulose column. This method was rapid, stable, versatile and appliable to high-throughput N-linked glycans production before ESI-MS analysis fromμg grade bio-samples glycoproteins and propitious to subsequent glycomic research.
本研究先以牛胰核糖核酸酶B和鸡卵清白蛋白二种常见的糖蛋白为研究材料,对几种N-糖链解离、富集纯化方法进行了考察,得到以下结论：
1.N-糖链解离方法：选用十二烷基磺酸钠(SDS)和碘乙酰胺(DTT)作蛋白质的变性剂,100℃反应10 min；加入磷酸钠缓冲液和10%乙基苯基聚乙二醇(NP40),然后加入N-糖苷酶F (PNGase F),37℃反应24 h。这种方法解离效果好,效率最高,操作简单。
2.N-糖链富集纯化方法：我们将富集纯化方法联合使用,对解离样品进行质谱分析前处理,结果表明,除了微晶纤维素和阳离子交换树脂混合装柱法外,其余4种方法(微晶纤维素柱结合阳离子交换树脂,C18柱结合活性炭柱,C18柱结合阳离子交换树脂和活性炭柱结合微晶纤维素柱)均可以达到满意的富集纯化效果。
我们将初步建立的方法用胎牛血清,健康小鼠血清,健康人血清,小鼠HAbI8 IgG以及HAbI8 IgG(腹水法)这5种生物样品进一步验证,并针对微量样品进行了改进：
3.PVDF膜辅助N-糖链解离方法：我们利用PVDF膜能够结合蛋白质的特性,在释放糖链前对复杂生物样品进行粗纯化,再按照上述糖链解离步骤,直接在膜上完成糖蛋白变性和糖链解离过程。实验证明,这种方法能同时提高微量样品糖链纯化效果。
4.改进的N-糖链富集纯化方法：上述4种富集纯化方法中,只有活性炭柱联合微晶纤维素柱纯化生物来源N-糖链解离样品的效果最好。在用活性炭柱进行纯化时,为了避免糖链洗脱不完全,我们用含0.1%三氟乙酸的50%的乙腈溶液为洗脱剂；微晶纤维素柱的平衡时间与填料多少成正比,平衡效果好坏直接影响纯化效果,而微量样品只需要少量的微晶纤维素,因此我们采用1000μL移液器标配枪头作一次性层析柱,可以大大减少干扰物,提高纯化效率。
综上所述,本研究在电喷雾质谱检测技术的基础上,建立了一个更适合于微量生物来源样品糖蛋白N-糖链解离,富集纯化的新方法——采用PVDF膜辅助N-糖链解离方法以及活性炭柱联合微晶纤维素柱的富集纯化方法。这种方法快速,稳定,通用性更强,能够通量化应用于ESI-MS检测前μg级别生物样品糖蛋白N-糖链的制备,有利于糖组的后续研究。
Glycobiology is an important part of life science research in 21st century. To have in-depth study of the functions and regulatory mechanism of glycans in life activities, the primary task is to obtain and analyze glycome. Electrospray Ionization Mass Spectrometry (ESI-MS) is a powerful tool to analyze micro-scale glycomes, but the pretreatment of sample is necessary. In this paper, we established a pretreatment procedure for N-linked glycans released from glycoproteins in micro-scale bio-samples before ESI-MS analysis in view of the exsting problems.
This study firstly used two common glycoproteins, bovine ribonuclease B and chicken albumin, as research materials, and had an initial investigation on several releasing, purifying and enriching N-linked glycans methods and obtained the following conclusions:
1. N-linked glycans releasing:we chose sodium dodecyl sulfate (SDS) and iodine acetamide (DTT) as protein denaturing agents. Glycoproteins were dissolved in the buffer and denatured at 100℃for 10 min. Then added sodium pyrophosphate buffer,10% ethyl-phenyl-polyethyleneglycol (NP40), PNGase F,37℃24 h. This approach is effective, efficient and easy to operate.
2. N-linked glycans purification and enrichment:the enzymatic samples were pretreated separately by five enrichment and purification methods used in conjunction before mass spectrometry analysis. The results showed, in addition to the method of using microcrystalline cellulose and cation exchange resin in tandem, the other four kinds (combination of microcrystalline cellulose and cation exchange resin column, combination of C18 column and graphite carbon column, combination of C18 column and cation exchange resin and combination of graphite carbon column and microcrystalline cellulose column) were able to achieve a satisfactory effect of purification and enrichment.
Then, we used five bio-samples, fetus bovine serum, control mouse serum, control human serum, mouse HAbI8 IgG and HAbI8 IgG (ascites method) for further verification. Moreover, we improved this method focusing on micro-scale samples:
3. PVDF membrane assisted N-linked glycans releasing:the PVDF membrane has the characteristic of absorbing proteins. We used it to roughly purify the complex bio-samples prior to PNGase F digestion, and then in accordance with the above steps completed glycoprotein denaturation and enzymatic digestion directly on the membrane. The results showed that this method can also improve the purification effect of glycans from micro-scale samples.
4. Improved procedure of purifying and enriching.N-linked glycans:In the above mentioned four kinds of enrichment and purification methods, only the combination of graphite carbon column and microcrystalline cellulose column had the best purifying and enriching effect. We used 0.1% trifluoroacetic acid containing 50% acetonitrile solution as eluting agent of graphite carbon column to avoid eluting free glycans incompelely; the pretreatment time of microcrystalline cellulose powder was related with quantities, and the effect of pretreatment would have a direct impact on glycans analysis. The micro-scale samples only required a small amount of powder, so we made use of the 1000μL standard pipettes as the disposable chromatography columns and greatly reduced impurities, improved the efficiency of purification.
This study, based on ESI-MS technology, established a new procedure which was more suitable for purifying and enriching N-linked glycans from micro-scale bio-samples glycoproteins--PVDF membrane assisted N-linked glycans releasing, purifying and enriching free glycans by graphite carbon column and microcrystalline cellulose column. This method was rapid, stable, versatile and appliable to high-throughput N-linked glycans production before ESI-MS analysis fromμg grade bio-samples glycoproteins and propitious to subsequent glycomic research.
引文
[1]Varki A. Biological roles of oligosaccharides:all of the theories are correct[J]. Glycobiology,1993,3 (2):97-130
[2]Krueger K. E., Srivastava S. Posttranslational Protein Modifications:Current Implications for Cancer Detection, Prevention, and Therapeutics[J]. Mol. Cell. Proteomics,2006,5 (10):1799-1810
[3]Drickamer K., Taylor M. E. Evolving views of protein glycosylation[J]. Trends Biochem. Sci.,1998,23 (9):321-324
[4]Dwek R. A. Glycobiology:towards understanding the function of sugars[J]. Biochem. Soc. Trans.,1995,23(1):1-25
[5]Lis H., Sharon N. Protein glycosylation. Structural and functional Aspects[J]. Eur. J. Biochem.,1993,218(1):1-27
[6]Lowe J. B. Glycan-dependent leukocyte adhesion and recruitment in inflammation[J]. Curr. Opin. Cell Biol.,2003,15 (5):531-538
[7]Fukuda M. Possible roles of tumor-associated carbohydrate antigens[J]. Cancer Res.1996, 56(10):2237-2244
[8]Lowe J. B., Marth J. D. A genetic approach to Mammalian glycan function[J]. Annu. Rev. Biochem.,2003,72:643-691
[9]张真庆.质谱分析外源性和内源性寡糖的方法学研究[D].青岛：中国海洋大学,2006
[10]Olden K. Adhesion molecules and inhibitors of glycosylation in cancer[J]. Semin. Cancer Biol.,1993,4 (5):269-276
[11]Hirabayashi J., Arata Y., Kasai. Glycome project:concept, strategy and preliminary application to Caenorhabditis elegans[J]. Proteomics,2001, 1(2):295-303
[12]Taniguchi N., Ekuni A., Ko J H, et al. A glycomic approach to the identification and characterization of glycoprotein function in cells transfected with glycosyltransferase genes[J]. Proteomics,2001, 1(2):239-247
[13]Pilobello K. T, Mahal L. K. Deciphering the glycocode:the complexity and analytical challenge of glycomics[J]. Current Opinion in Chemical Biology,2007,11:300-305
[14]Kam R. K. T.; Poon, T. C. W. The Potentials of Glycomics in Biomarker Discovery[J]. Clinical Proteomics,2008,4:67-79
[15]Haslam S. M., North S. J., Dell A. Mass spectrometric analysis of N-and O-glycosylation of tissues and cells[J]. Current Opinion in Structural Biology,2006, 16:584-591
[16]Taniguchi N., Honke K., Fukuda M. Handbook of Glycosyltransferases and Related Genes[M]. Springer,2002:1-2
[17]Dwek, M.V. and Brooks, S.A. Harnessing changes in cellular glycosylation in new cancer treatment strategies. Curr. Cancer Drug Targets,2004,(4):425-442
[18]莫琳·E·泰勒,库尔特·德里卡默.糖生物学导论[M].北京：化学工业出版社,2006：26-100
[19]Lowe J.B. Glycosylation, Immunity, and Autoimmunity[J]. Cell,2001,104(6):809-812
[20]Ohtsubo Kazuaki, Marth Jamey D. Glycosylation in Cellular Mechanisms of Health and Disease[J]. Cell,2006,126:855-867
[21]沈同,王镜岩,等.生物化学[M].第二版.北京：高等教育出版社,1990：57-154
[22]夏其昌,曾嵘.蛋白质化学与蛋白质组学[M].北京：科学出版社,2004：374-393
[23]Ritchie M. A., Gill A. C., Deery M. J. Precursor Ion Scanning for Detection and Structural Characterization of Heterogeneous Glycopeptide Mixtures[J]. American Society for Mass Spectrometry,2002,13:1065-1077
[24]杨珺,蔡邵皙,邹全明.糖组学研究技术及其进展[J].生物化学与生物物理进展,2005,32(1)：9-12
[25]Rademacher T.W., Parekh R.B., Dwek R.A. Glycobiology[J]. Annu. Rev. Biochem,1988, 57:785-838
[26]Furukawa K., Kobata A. Protein glycosylation[J]. Curr. Opin. Biotechnol.,1992, 3:554-559
[27]Charlwood J., Birrell H., Camilleri P. Carbohydrate release from picomole quantities of glycoprotein and characterisation of glycans by high-performance liquid chromatography and mass spectrometry[J]. Phytochemistry,1999,51(2):199-210
[28]Miyako Nakano, Akihiro Kondo, Kazuaki Kakehi, et al. Glycomics-a new target for pharmaceuticals[J]. Analytical chemistry,2006,3(1):39-47
[29]王克夷,杜昱光.糖组和糖组学[J].医学分子生物学杂志,2004,1(5)：261-264
[30]Varki A., Cummings R., Esko J., et al. Essentials of glycobiology[M]. Cold Spring Harbor Laboratory,1999:537-540
[31]An Hyun Joo, Miyamoto Suzanne, Lancaster Katherine S., et al. Profiling of Glycans in Serum for the Discovery of Potential Biomarkers for Ovarian Cancer[J]. Journal of Proteome Research,2006, (5):1626-1635
[32]Saldova R., Royle L., Radcliffe C.M., et al. Ovarian Cancer is Associated with Changes in Glycosylation in Both Acute-Phase Proteins and IgG[J]. Glycobiology,2007,17(12): 1344-1356
[33]Saldova R., Wormald M. R., Dwek R. A., et al. Glycosylation changes on serum glycoproteins in ovarian cancer may contribute to disease pathogenesis[J]. Disease Markers,2008,25:219-232
[34]Okuyama N., Ide Y., Nakano M., et al. Fucosylated haptoglobin is a novel marker for pancreatic cancer:A detailed analysis of the oligosaccharide structure and a possible mechanism for fucosylation[J]. Int. J. Cancer,2006,118 (11):2803-2808
[35]Otake Y., Fujimoto I., Tanaka F., et al. Isolation and Characterization of an N-Linked Oligosaccharide That Is Significantly Increased in Sera from Patients with Non-Small Cell Lung Cancer[J]. J. Biochem.,2001,129 (4):537-542.
[36]Kossowska B., Ferens-Sieczkowska M., Gancarz R., et al. Fucosylation of serum glycoproteins in lung cancer patients [J]. Clin. Chem. Lab. Med.,2005,43:361-369
[37]Comunale M.A., Lowman M., Long R.E., et al. Proteomic Analysis of Serum Associated Fucosylated Glycoproteins in the Development of Primary Hepatocellular Carcinoma[J]. Journal of Proteome Research,2006, (5):308-315
[38]Isailovic D., Kurulugama R. T., Plasencia M. D., et al. Profiling of Human Serum Glycans Associated with Liver Cancer and Cirrhosis by IMS-MS [J]. Journal of Proteome Research,2008, (7):1109-1117
[39]Toppila S., Paavonen T., Laitinen A., et al. Endothelial sulfated sialyl Lewis X glycans, putative L-selectin ligands, are preferentially expressed in bronchial asthma but not in other chronic inflammatory lung diseases[J]. Am. J. Respir. Cell Mol. Biol.,2000,23: 492-498
[40]Fernandes B., Sagman U., Auger M., et al. Beta 1-6 branched oligosaccharides as a marker of tumor progression in human breast and colon neoplasia[J]. Cancer Res.,1991, 51:718-723
[41]Taniguchi N., Miyoshi E., Ko J.H.,et al. Implication of N-acetylglucosaminyl transferases III and V in cancer:gene regulation and signaling mechanism[J]. Biochim., Biophys. Acta,1999,1455:287-300
[42]谢红东,周滔,刘成海.肝脏疾病蛋白质组学研究进展[J].中西医结合肝病杂志,2007,17(3)：183-185
[43]彭小宁,彭司华,曾小敏.卵巢癌早期诊断方法的研究进展[J].国际病理科学与临床杂志,2007,27(6)：489-492
[44]Gloeckler Ries L.A., Reichman M. E., Lewis D. R., et al. Cancer survival and incidence, from the surveillance, epidemiology, and end results (SEER) program[J]. Oncologist, 2003, (8):541-552
[45]Semmes O.J., Feng Z., Adam B.L., et al. Evaluation of serum protein profiling by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry for the detection of prostate cancer:I. Assessment of platform reproducibility[J]. Clin. Chem., 2005,51:102-112
[46]聂赣娟,周建华,李茂玉等.肺鳞癌患者与健康人血清的差异蛋白质组学研究[J].生物化学与生物物理进展,2008,35(3)：349-355
[47]邱惠麒,张稚琴.肿瘤标志物CA125在妇科疾病中的应用[J].浙江预防医学,1999,(8)：30-31
[48]Sonoda K., Miyamoto S., Hirakawa Toshio, et al. Clinical significance of RCAS1 as a biomarker of uterine cancer[J]. Gynecologic Oncology,2006,103:924-931
[49]Dennis J. W., Granovsky M., Warren C. E. Glycoprotein glycosylation and cancer progression[J]. Biochin. Biophys. Acta.,1999,1473:21-34
[50]Dube D. H., Bertozzi C. R. Glycans in cancer and inflammation-potential for therapeutics and diagnostics[J]. Nat. Rev. Drug Discovery,2005, (4):477-488
[51]Gornik I., Maravic G., Dumic J., et al. Fucosylation of IgG Heavy Chains is Increased in Rheumatoid Arthritis[J]. Clinical Biochemistry,1999,32(8):605-608
[52]Taniguchi N., Hancock W., Lubman D. M., et al.The Second Golden Age of Glycomics: From Functional Glycomics to Clinical Applications[J]. J. Proteome Res.,2009,8 (2): 425-426
[53]卢雁,李向荣.蛋白质变性机理与变性时的热力学参数研究进展[J].化学进展,2005,(5)：906-910
[54]杜厚三.浅谈蛋白质的变性作用[J].食品科技,1983,(3)：21
[55]杨平,李敏惠,常山等.二硫苏糖醇和β-巯基乙醇在SDS-PAGE电泳过程中的扩散效应[J].医学研究生学报,2009,(9)：916-919
[56]刘静,徐桂英.表面活性剂与蛋白质相互作用的研究进展[J].日用化学工业,2003,(1)：29-32
[57]Liu X., Li X.Y., Chan K., et al. "One-Pot" Methylation in Glycomics Application: Esterification of Sialic Acids and Permanent Charge Construction[J]. Analytical Chemistry,2007,79:3894-3900
[58]Tarentino A.L., Gomez C.M., Plummer Jr.T.H. Deglycosylation of asparagine-linked glycans by peptide:N-glycosidase F[J]. Biochemistry,1985, (24):4665
[59]Lee J. Y, Park T. H. Enzyme and Microbial Technology [J],2002,30(6):716-720
[60]Tretter V., Altmann F., Marz L.Peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F cannot release glycans with fucose attached alpha 1-3 to the asparagine-linked N-acetylglucosamine residue[J]. Eur. J. Biochem,1991,199(3):647-652
[61]Tarentio A. L., Plumer T. H. Enzymatic deglycosylation of asparagine-linked glycans: Purification, properties, and specificity of oligosaccharide-cleaving enzymes from Flavobacterium meningosepticum[J]. Meth. Enzymol.,1994,230:44-57
[62]Tarentino A. L., Plummer T. H. Jr., Maley F. The Release of Intact Oligosaccharides from Specific Glycoproteins by Endo-β-N-acetylglucosaminidase H[J]. The Journal of Biological Chemistry,1974,249:818-824
[63]Takasaki S. Mizuochi T., Kobata A.. Hydrazinolysis of asparagine-linked sugar chains to produce free oligosaccharides[J]. Methods in Enzymology,1982,83:263-268
[64]Patel T., Bruce J., Merry A., et al. Use of Hydrazine To Release in Intact and Unreduced Form both N-and O-Linked Oligosaccharides from Glycoproteins[J]. Biochemistry,1993, 32:679-693
[65]苏立强,郑永杰.色谱分析法[M].北京：清华大学出版社,2009：385-394
[66]刘文韬,刘炳亚,朱正纲.糖蛋白质组学研究技术[J].诊断学理论与实践,2006, 5(4)：365-367
[67]王川.糖组学：破解生命信息的第3种途径[J].生物学通报,2005,40(5)：8-9
[68]Satish P.R., Surolia A. Exploiting lectin affinity chromatography in clinical diagnosis[J]. J Biochem Biophys Methods,2001,49:625-640
[69]Nakagawa H., Hato M., Takegawa Y., et al. Detection of altered N-glycan profiles in whole serum from rheumatoid arthritis patients[J]. Journal of Chromatography B,2007, 853:133-137
[70]王晨,田庚元.糖蛋白中糖链的结构研究[J].化学通报,1994,(5)：8-11
[71]于泓,牟世芬.离子色谱中的安培检测方法及其应用[J].化学通报,2007,70(7)：1-10
[72]牟世芬,于泓,蔡亚岐.糖的高效阴离子交换色谱-脉冲安培检测法分析[J].色谱,2009,27(5)：667-674
[73]Zuzana Kyselova, Yehia Mechref, Pilsoo Kang, et al. Breast Cancer Diagnosis and Prognosis through Quantitative Measurements of Serum Glycan Profiles[J]. Clinical Chemistry,2008,54(7):1166-1175
[74]Fauquenoy S., Morelle W., Hovasse A., et al. Proteomics and Glycomics Analyses of N-Glycosylated Structures Involved in Toxoplasma gondii-Host Cell Interactions[J]. Molecular & Cellular Proteomics,2008, (7):891-910
[75]袁黎明.制备色谱技术及应用[M].北京：化学工业出版社,2005：38-88
[76]Packer N.H., Lawson M.A., Jardine D.R., et al. A general approach to desalting oligosaccharides released from glycoproteins[J]. Glycoconjugate Journal,1998, 15:737-747
[77]Michael G., Novalin S., Wendland M., et al. Desalination of whey by electrodialysis and ion exchange resins:analysis of both processes with regard to sustainability by calculating their cumulative energy demand[J]. Journal of Membrane Science,2002,210:91-102
[78]Plon Ka am. Characteristics of Microcrystalline and Microfine Cellulose[J]. Cellul Chem Technol.,1982,16:473-483
[79]Shimizu Y, Nakata M., Kuroda Y, et al. Rapid and simple preparation of N-linked oligosaccharides by cellulose-column chromatography [J]. Carbohydrate Research,2001, 332:381-388
[80]Ashline D. J., Lapadula A. J., Liu Y. H., et al. Carbohydrate Structural Isomers Analyzed by Sequential Mass Spectrometry[J]. Analytical Chemistry,2007,79:3830-3842
[81]Xia B. Y., Feasley C.L., Sachdev G.P., et al. Glycan reductive isotope labeling for quantitative glycomics[J].Anal. Biochem.,2009,387:162-170
[82]陈耀祖,涂亚平.有机质谱原理及应用[M].北京：科学出版社,2001：2-15
[83]何美玉.现代有机与生物质谱[M].北京：北京大学出版社,2002：3-17
[84]汪尔康.21世纪的分析化学[M].北京：科学出版社,2001：297
[85]刘翠平,方积年.质谱技术在糖类结构分析中的应用[J].分析化学评述与进展,2001,29(6)：716-720
[86]Weiskopf A. S., Vouros P., Harvey D.J. Electrospray Ionization-Ion Trap Mass Spectrometry for Structural Analysis of Complex N-Linked Glycoprotein Oligosaccharides[J]. Anal. Chem.,1998,70:4441-4447
[87]Elaine Stephens, Sarah L. Maslen, Luke G Green, Dudley H. Williams, Fragmentation Characteristics of Neutral N-Linked Glycans Using a MALDI-TOF/TOF Tandem Mass Spectrometer, Anal. Chem.2004,76:2343-2354
[88]Wilm M., Mann M. Analytical Properties of the Nanoelectrospray Ion Source[J]. Anal. Chem.,1996,68 (1):1-8
[89]Valaskovic G.A., Kelleher N.L., Little D.P., et al. Attomole-Sensitivity Electrospray Source for Large-Molecule Mass Spectrometry[J]. Anal. Chem.,1995,67 (20): 3802-3805
[90]Julianne C. B., James A. A. Ⅲ, Lei C., et al. Quantification by isobaric labeling (QUIBL) for the comparative glycomic study of O-linked glycans[J]. Int. J. Mass Spectrom.,2008, 278:137-142
[91]Kita Y., Miura Y., Furukawa J., et al. Quantitative Glycomics of Human Whole Serum Glycoproteins Based on the Standardized Protocol for Liberating N-Glycans[J]. Molecular & Cellular Proteomics,2007,6:1437-1445
[92]卢庄,王京兰,蔡耘等.糖蛋白组学研究技术进展及其应用[J].生命的化学,2008,28(3)：335-338
[93]Tang L., Kebarle P. Effect of the conductivity of the electrosprayed solution on the electrospray current. Factors determining analyte sensitivity in electrospray mass spectrometry[J]. Analytical chemistry,1991,63:2709-2715
[94]Harazonoa A., Kawasakia N., Itoh S., et al. Simultaneous glycosylation analysis of human serum glycoproteins by high-performance liquid chromatography/tandem mass spectrometry[J]. Journal of Chromatography B,2008,869:20-30
[95]Tzur Yaron, Markovich Amir, Lichtenstein Rachel G. A Two-Dimensional Array for Simultaneous Sequencing of N-and O-Glycans and Their Glycoforms on Specific Glycosylation Sites[J]. Journal of Proteome Research,2008,7:1188-1198
[96]Kang Pilsoo, Mechref Yehia, Novotny Milos V. High-throughput solid-phase permethylation of glycans prior to mass spectrometry[J]. Rapid Communication in Mass Spectrometry,2008,22:721-734
[97]Suzuki S., Tanaka R., Takada K., et al. Analysis of sialo-N-glycans in glycoproteins as 1-phenyl-3-methyl-5-pyrazolone derivatives by capillary electrophoresis[J]. Journal of Chromatography A,2001,910:319-329
[98]Marcell O., Peter H., Guenther K. B., et al. Sample Preparation for the Analysis of Complex Carbohydrates by Multicapillary Gel Electrophoresis with Light-Emitting Diode Induced Fluorescence Detection[J]. Analytical Chemistry,2008,80:4241-4246
[99]Kyselova Z., Mechref Y., Al Bataineh M.M., et al. Alterations in the serum glycome due to metastatic prostate cancer[J]. J. Proteome Res.,2007,6:1822-1832
[100]Gil G. C., Kim Y. G., Kim B. G. A relative and absolute quantification of neutral N-linked oligosaccharides using modification with carboxymethyl trimethylammonium hydrazide and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry[J]. Analytical Biochemistry,2008,379:45-59
[101]Kang P., Mechref Y., Kyselova Z., et al. Comparative Glycomic Mapping through Quantitative Permethylation and Stable-Isotope Labeling[J]. Analytical Chemistry,2007, 79:6064-6073
[102]Weiskopf A.S., Vouros P., Harvey D.J. Characterization of oligosaccharide composition and structure by quadrupole ion trap mass spectrometry[J]. Rapid. Commun. Mass Spectrom.,1997,11:1493-1504
[103]Huntington J.A., Stein P.E. Structure and properties of ovalbumin[J]. Journal of Chromatography B,2001,756:189-198
[104]Harvey D. J. Postsource decay fragmentation of N-linked carbohydrates from ovalbumin and related glycoproteins[J]. American Society for Mass Spectrometry.2000, 11:572-577
[105]Harvey D. J., Wing D. R., Kuster B., et al. Composition of N-linked carbohydrates from ovalbumin and co-purified glycoproteins[J]. American Society for Mass Spectrometry, 2000,11:564-571
[106]廖秋林,陈晓东,丁彦青.血清蛋白质组学技术及研究进展[J].中国病理生理杂志,2008,24(2)：402-407
[107]Royle L., Campbell M. P., Radcliffe C. M., et al. Rapid and high-throughput analysis of N-glycans from ovarian cancer serum using a 96-well plate platform[J]. Analytical Biochemistry,2009,391:1-12
[108]周军,刘云叶,长明等.聚偏氟乙烯(PVDF)膜的制备及在水处理中应用的研究[J].新技术新产品,2007,(12)：51-55
[109]Masaaki Toyoda, Hiromi Ito, Yu-ki Matsuno, et al. Quantitative Derivatization of Sialic Acids for the Detection of Sialoglycans by MALDI MS[J]. Anal. Chem.,2008,80: 5211-5218
[110]Geoffrey R. Guile, Pauline M. Rudd, David R. Wing, et al. A Rapid High-Resolution High-Performance Liquid Chromatographic Method for Separating Glycan Mixtures and Analyzing Oligosaccharide Profiles[J], Analytical biochemistry,1996,240:210-226
[111]Lattova E., Varma S.,Bezabeh T., et al. Mass spectrometric profiling of N-Linked oligosaccharides and uncommon glycoform in mouse serum with head and neck tumor[J]. J Am Soc Mass Spectrom,2008,19:671-685
[112]Atwood J. A. Ⅲ, Cheng L., Alvarez-Manilla G., et al. Quantitation by isobaric labeling: applications to glycomics[J]. Journal of Proteome Research,2008, (7):367-374
[113]Harvey D. J., Royle L., Radcliffe C.M., et al. Structural and quantitative analysis of N-linked glycans by matrix-assisted laser desorption ionization and negative ion nanospray mass spectrometry[J], Analytical Biochemistry,2008,376:44-60
[114]Morten T. A., Simon M., Peter H. Site-Specific Glycoprofiling of N-Linked glycopeptides Using MALDI-TOF MS:strong correlation between signal strength and glycoform quantities[J]. Anal. Chem.,2009,81:3933-3943
[2]Krueger K. E., Srivastava S. Posttranslational Protein Modifications:Current Implications for Cancer Detection, Prevention, and Therapeutics[J]. Mol. Cell. Proteomics,2006,5 (10):1799-1810
[3]Drickamer K., Taylor M. E. Evolving views of protein glycosylation[J]. Trends Biochem. Sci.,1998,23 (9):321-324
[4]Dwek R. A. Glycobiology:towards understanding the function of sugars[J]. Biochem. Soc. Trans.,1995,23(1):1-25
[5]Lis H., Sharon N. Protein glycosylation. Structural and functional Aspects[J]. Eur. J. Biochem.,1993,218(1):1-27
[6]Lowe J. B. Glycan-dependent leukocyte adhesion and recruitment in inflammation[J]. Curr. Opin. Cell Biol.,2003,15 (5):531-538
[7]Fukuda M. Possible roles of tumor-associated carbohydrate antigens[J]. Cancer Res.1996, 56(10):2237-2244
[8]Lowe J. B., Marth J. D. A genetic approach to Mammalian glycan function[J]. Annu. Rev. Biochem.,2003,72:643-691
[9]张真庆.质谱分析外源性和内源性寡糖的方法学研究[D].青岛：中国海洋大学,2006
[10]Olden K. Adhesion molecules and inhibitors of glycosylation in cancer[J]. Semin. Cancer Biol.,1993,4 (5):269-276
[11]Hirabayashi J., Arata Y., Kasai. Glycome project:concept, strategy and preliminary application to Caenorhabditis elegans[J]. Proteomics,2001, 1(2):295-303
[12]Taniguchi N., Ekuni A., Ko J H, et al. A glycomic approach to the identification and characterization of glycoprotein function in cells transfected with glycosyltransferase genes[J]. Proteomics,2001, 1(2):239-247
[13]Pilobello K. T, Mahal L. K. Deciphering the glycocode:the complexity and analytical challenge of glycomics[J]. Current Opinion in Chemical Biology,2007,11:300-305
[14]Kam R. K. T.; Poon, T. C. W. The Potentials of Glycomics in Biomarker Discovery[J]. Clinical Proteomics,2008,4:67-79
[15]Haslam S. M., North S. J., Dell A. Mass spectrometric analysis of N-and O-glycosylation of tissues and cells[J]. Current Opinion in Structural Biology,2006, 16:584-591
[16]Taniguchi N., Honke K., Fukuda M. Handbook of Glycosyltransferases and Related Genes[M]. Springer,2002:1-2
[17]Dwek, M.V. and Brooks, S.A. Harnessing changes in cellular glycosylation in new cancer treatment strategies. Curr. Cancer Drug Targets,2004,(4):425-442
[18]莫琳·E·泰勒,库尔特·德里卡默.糖生物学导论[M].北京：化学工业出版社,2006：26-100
[19]Lowe J.B. Glycosylation, Immunity, and Autoimmunity[J]. Cell,2001,104(6):809-812
[20]Ohtsubo Kazuaki, Marth Jamey D. Glycosylation in Cellular Mechanisms of Health and Disease[J]. Cell,2006,126:855-867
[21]沈同,王镜岩,等.生物化学[M].第二版.北京：高等教育出版社,1990：57-154
[22]夏其昌,曾嵘.蛋白质化学与蛋白质组学[M].北京：科学出版社,2004：374-393
[23]Ritchie M. A., Gill A. C., Deery M. J. Precursor Ion Scanning for Detection and Structural Characterization of Heterogeneous Glycopeptide Mixtures[J]. American Society for Mass Spectrometry,2002,13:1065-1077
[24]杨珺,蔡邵皙,邹全明.糖组学研究技术及其进展[J].生物化学与生物物理进展,2005,32(1)：9-12
[25]Rademacher T.W., Parekh R.B., Dwek R.A. Glycobiology[J]. Annu. Rev. Biochem,1988, 57:785-838
[26]Furukawa K., Kobata A. Protein glycosylation[J]. Curr. Opin. Biotechnol.,1992, 3:554-559
[27]Charlwood J., Birrell H., Camilleri P. Carbohydrate release from picomole quantities of glycoprotein and characterisation of glycans by high-performance liquid chromatography and mass spectrometry[J]. Phytochemistry,1999,51(2):199-210
[28]Miyako Nakano, Akihiro Kondo, Kazuaki Kakehi, et al. Glycomics-a new target for pharmaceuticals[J]. Analytical chemistry,2006,3(1):39-47
[29]王克夷,杜昱光.糖组和糖组学[J].医学分子生物学杂志,2004,1(5)：261-264
[30]Varki A., Cummings R., Esko J., et al. Essentials of glycobiology[M]. Cold Spring Harbor Laboratory,1999:537-540
[31]An Hyun Joo, Miyamoto Suzanne, Lancaster Katherine S., et al. Profiling of Glycans in Serum for the Discovery of Potential Biomarkers for Ovarian Cancer[J]. Journal of Proteome Research,2006, (5):1626-1635
[32]Saldova R., Royle L., Radcliffe C.M., et al. Ovarian Cancer is Associated with Changes in Glycosylation in Both Acute-Phase Proteins and IgG[J]. Glycobiology,2007,17(12): 1344-1356
[33]Saldova R., Wormald M. R., Dwek R. A., et al. Glycosylation changes on serum glycoproteins in ovarian cancer may contribute to disease pathogenesis[J]. Disease Markers,2008,25:219-232
[34]Okuyama N., Ide Y., Nakano M., et al. Fucosylated haptoglobin is a novel marker for pancreatic cancer:A detailed analysis of the oligosaccharide structure and a possible mechanism for fucosylation[J]. Int. J. Cancer,2006,118 (11):2803-2808
[35]Otake Y., Fujimoto I., Tanaka F., et al. Isolation and Characterization of an N-Linked Oligosaccharide That Is Significantly Increased in Sera from Patients with Non-Small Cell Lung Cancer[J]. J. Biochem.,2001,129 (4):537-542.
[36]Kossowska B., Ferens-Sieczkowska M., Gancarz R., et al. Fucosylation of serum glycoproteins in lung cancer patients [J]. Clin. Chem. Lab. Med.,2005,43:361-369
[37]Comunale M.A., Lowman M., Long R.E., et al. Proteomic Analysis of Serum Associated Fucosylated Glycoproteins in the Development of Primary Hepatocellular Carcinoma[J]. Journal of Proteome Research,2006, (5):308-315
[38]Isailovic D., Kurulugama R. T., Plasencia M. D., et al. Profiling of Human Serum Glycans Associated with Liver Cancer and Cirrhosis by IMS-MS [J]. Journal of Proteome Research,2008, (7):1109-1117
[39]Toppila S., Paavonen T., Laitinen A., et al. Endothelial sulfated sialyl Lewis X glycans, putative L-selectin ligands, are preferentially expressed in bronchial asthma but not in other chronic inflammatory lung diseases[J]. Am. J. Respir. Cell Mol. Biol.,2000,23: 492-498
[40]Fernandes B., Sagman U., Auger M., et al. Beta 1-6 branched oligosaccharides as a marker of tumor progression in human breast and colon neoplasia[J]. Cancer Res.,1991, 51:718-723
[41]Taniguchi N., Miyoshi E., Ko J.H.,et al. Implication of N-acetylglucosaminyl transferases III and V in cancer:gene regulation and signaling mechanism[J]. Biochim., Biophys. Acta,1999,1455:287-300
[42]谢红东,周滔,刘成海.肝脏疾病蛋白质组学研究进展[J].中西医结合肝病杂志,2007,17(3)：183-185
[43]彭小宁,彭司华,曾小敏.卵巢癌早期诊断方法的研究进展[J].国际病理科学与临床杂志,2007,27(6)：489-492
[44]Gloeckler Ries L.A., Reichman M. E., Lewis D. R., et al. Cancer survival and incidence, from the surveillance, epidemiology, and end results (SEER) program[J]. Oncologist, 2003, (8):541-552
[45]Semmes O.J., Feng Z., Adam B.L., et al. Evaluation of serum protein profiling by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry for the detection of prostate cancer:I. Assessment of platform reproducibility[J]. Clin. Chem., 2005,51:102-112
[46]聂赣娟,周建华,李茂玉等.肺鳞癌患者与健康人血清的差异蛋白质组学研究[J].生物化学与生物物理进展,2008,35(3)：349-355
[47]邱惠麒,张稚琴.肿瘤标志物CA125在妇科疾病中的应用[J].浙江预防医学,1999,(8)：30-31
[48]Sonoda K., Miyamoto S., Hirakawa Toshio, et al. Clinical significance of RCAS1 as a biomarker of uterine cancer[J]. Gynecologic Oncology,2006,103:924-931
[49]Dennis J. W., Granovsky M., Warren C. E. Glycoprotein glycosylation and cancer progression[J]. Biochin. Biophys. Acta.,1999,1473:21-34
[50]Dube D. H., Bertozzi C. R. Glycans in cancer and inflammation-potential for therapeutics and diagnostics[J]. Nat. Rev. Drug Discovery,2005, (4):477-488
[51]Gornik I., Maravic G., Dumic J., et al. Fucosylation of IgG Heavy Chains is Increased in Rheumatoid Arthritis[J]. Clinical Biochemistry,1999,32(8):605-608
[52]Taniguchi N., Hancock W., Lubman D. M., et al.The Second Golden Age of Glycomics: From Functional Glycomics to Clinical Applications[J]. J. Proteome Res.,2009,8 (2): 425-426
[53]卢雁,李向荣.蛋白质变性机理与变性时的热力学参数研究进展[J].化学进展,2005,(5)：906-910
[54]杜厚三.浅谈蛋白质的变性作用[J].食品科技,1983,(3)：21
[55]杨平,李敏惠,常山等.二硫苏糖醇和β-巯基乙醇在SDS-PAGE电泳过程中的扩散效应[J].医学研究生学报,2009,(9)：916-919
[56]刘静,徐桂英.表面活性剂与蛋白质相互作用的研究进展[J].日用化学工业,2003,(1)：29-32
[57]Liu X., Li X.Y., Chan K., et al. "One-Pot" Methylation in Glycomics Application: Esterification of Sialic Acids and Permanent Charge Construction[J]. Analytical Chemistry,2007,79:3894-3900
[58]Tarentino A.L., Gomez C.M., Plummer Jr.T.H. Deglycosylation of asparagine-linked glycans by peptide:N-glycosidase F[J]. Biochemistry,1985, (24):4665
[59]Lee J. Y, Park T. H. Enzyme and Microbial Technology [J],2002,30(6):716-720
[60]Tretter V., Altmann F., Marz L.Peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F cannot release glycans with fucose attached alpha 1-3 to the asparagine-linked N-acetylglucosamine residue[J]. Eur. J. Biochem,1991,199(3):647-652
[61]Tarentio A. L., Plumer T. H. Enzymatic deglycosylation of asparagine-linked glycans: Purification, properties, and specificity of oligosaccharide-cleaving enzymes from Flavobacterium meningosepticum[J]. Meth. Enzymol.,1994,230:44-57
[62]Tarentino A. L., Plummer T. H. Jr., Maley F. The Release of Intact Oligosaccharides from Specific Glycoproteins by Endo-β-N-acetylglucosaminidase H[J]. The Journal of Biological Chemistry,1974,249:818-824
[63]Takasaki S. Mizuochi T., Kobata A.. Hydrazinolysis of asparagine-linked sugar chains to produce free oligosaccharides[J]. Methods in Enzymology,1982,83:263-268
[64]Patel T., Bruce J., Merry A., et al. Use of Hydrazine To Release in Intact and Unreduced Form both N-and O-Linked Oligosaccharides from Glycoproteins[J]. Biochemistry,1993, 32:679-693
[65]苏立强,郑永杰.色谱分析法[M].北京：清华大学出版社,2009：385-394
[66]刘文韬,刘炳亚,朱正纲.糖蛋白质组学研究技术[J].诊断学理论与实践,2006, 5(4)：365-367
[67]王川.糖组学：破解生命信息的第3种途径[J].生物学通报,2005,40(5)：8-9
[68]Satish P.R., Surolia A. Exploiting lectin affinity chromatography in clinical diagnosis[J]. J Biochem Biophys Methods,2001,49:625-640
[69]Nakagawa H., Hato M., Takegawa Y., et al. Detection of altered N-glycan profiles in whole serum from rheumatoid arthritis patients[J]. Journal of Chromatography B,2007, 853:133-137
[70]王晨,田庚元.糖蛋白中糖链的结构研究[J].化学通报,1994,(5)：8-11
[71]于泓,牟世芬.离子色谱中的安培检测方法及其应用[J].化学通报,2007,70(7)：1-10
[72]牟世芬,于泓,蔡亚岐.糖的高效阴离子交换色谱-脉冲安培检测法分析[J].色谱,2009,27(5)：667-674
[73]Zuzana Kyselova, Yehia Mechref, Pilsoo Kang, et al. Breast Cancer Diagnosis and Prognosis through Quantitative Measurements of Serum Glycan Profiles[J]. Clinical Chemistry,2008,54(7):1166-1175
[74]Fauquenoy S., Morelle W., Hovasse A., et al. Proteomics and Glycomics Analyses of N-Glycosylated Structures Involved in Toxoplasma gondii-Host Cell Interactions[J]. Molecular & Cellular Proteomics,2008, (7):891-910
[75]袁黎明.制备色谱技术及应用[M].北京：化学工业出版社,2005：38-88
[76]Packer N.H., Lawson M.A., Jardine D.R., et al. A general approach to desalting oligosaccharides released from glycoproteins[J]. Glycoconjugate Journal,1998, 15:737-747
[77]Michael G., Novalin S., Wendland M., et al. Desalination of whey by electrodialysis and ion exchange resins:analysis of both processes with regard to sustainability by calculating their cumulative energy demand[J]. Journal of Membrane Science,2002,210:91-102
[78]Plon Ka am. Characteristics of Microcrystalline and Microfine Cellulose[J]. Cellul Chem Technol.,1982,16:473-483
[79]Shimizu Y, Nakata M., Kuroda Y, et al. Rapid and simple preparation of N-linked oligosaccharides by cellulose-column chromatography [J]. Carbohydrate Research,2001, 332:381-388
[80]Ashline D. J., Lapadula A. J., Liu Y. H., et al. Carbohydrate Structural Isomers Analyzed by Sequential Mass Spectrometry[J]. Analytical Chemistry,2007,79:3830-3842
[81]Xia B. Y., Feasley C.L., Sachdev G.P., et al. Glycan reductive isotope labeling for quantitative glycomics[J].Anal. Biochem.,2009,387:162-170
[82]陈耀祖,涂亚平.有机质谱原理及应用[M].北京：科学出版社,2001：2-15
[83]何美玉.现代有机与生物质谱[M].北京：北京大学出版社,2002：3-17
[84]汪尔康.21世纪的分析化学[M].北京：科学出版社,2001：297
[85]刘翠平,方积年.质谱技术在糖类结构分析中的应用[J].分析化学评述与进展,2001,29(6)：716-720
[86]Weiskopf A. S., Vouros P., Harvey D.J. Electrospray Ionization-Ion Trap Mass Spectrometry for Structural Analysis of Complex N-Linked Glycoprotein Oligosaccharides[J]. Anal. Chem.,1998,70:4441-4447
[87]Elaine Stephens, Sarah L. Maslen, Luke G Green, Dudley H. Williams, Fragmentation Characteristics of Neutral N-Linked Glycans Using a MALDI-TOF/TOF Tandem Mass Spectrometer, Anal. Chem.2004,76:2343-2354
[88]Wilm M., Mann M. Analytical Properties of the Nanoelectrospray Ion Source[J]. Anal. Chem.,1996,68 (1):1-8
[89]Valaskovic G.A., Kelleher N.L., Little D.P., et al. Attomole-Sensitivity Electrospray Source for Large-Molecule Mass Spectrometry[J]. Anal. Chem.,1995,67 (20): 3802-3805
[90]Julianne C. B., James A. A. Ⅲ, Lei C., et al. Quantification by isobaric labeling (QUIBL) for the comparative glycomic study of O-linked glycans[J]. Int. J. Mass Spectrom.,2008, 278:137-142
[91]Kita Y., Miura Y., Furukawa J., et al. Quantitative Glycomics of Human Whole Serum Glycoproteins Based on the Standardized Protocol for Liberating N-Glycans[J]. Molecular & Cellular Proteomics,2007,6:1437-1445
[92]卢庄,王京兰,蔡耘等.糖蛋白组学研究技术进展及其应用[J].生命的化学,2008,28(3)：335-338
[93]Tang L., Kebarle P. Effect of the conductivity of the electrosprayed solution on the electrospray current. Factors determining analyte sensitivity in electrospray mass spectrometry[J]. Analytical chemistry,1991,63:2709-2715
[94]Harazonoa A., Kawasakia N., Itoh S., et al. Simultaneous glycosylation analysis of human serum glycoproteins by high-performance liquid chromatography/tandem mass spectrometry[J]. Journal of Chromatography B,2008,869:20-30
[95]Tzur Yaron, Markovich Amir, Lichtenstein Rachel G. A Two-Dimensional Array for Simultaneous Sequencing of N-and O-Glycans and Their Glycoforms on Specific Glycosylation Sites[J]. Journal of Proteome Research,2008,7:1188-1198
[96]Kang Pilsoo, Mechref Yehia, Novotny Milos V. High-throughput solid-phase permethylation of glycans prior to mass spectrometry[J]. Rapid Communication in Mass Spectrometry,2008,22:721-734
[97]Suzuki S., Tanaka R., Takada K., et al. Analysis of sialo-N-glycans in glycoproteins as 1-phenyl-3-methyl-5-pyrazolone derivatives by capillary electrophoresis[J]. Journal of Chromatography A,2001,910:319-329
[98]Marcell O., Peter H., Guenther K. B., et al. Sample Preparation for the Analysis of Complex Carbohydrates by Multicapillary Gel Electrophoresis with Light-Emitting Diode Induced Fluorescence Detection[J]. Analytical Chemistry,2008,80:4241-4246
[99]Kyselova Z., Mechref Y., Al Bataineh M.M., et al. Alterations in the serum glycome due to metastatic prostate cancer[J]. J. Proteome Res.,2007,6:1822-1832
[100]Gil G. C., Kim Y. G., Kim B. G. A relative and absolute quantification of neutral N-linked oligosaccharides using modification with carboxymethyl trimethylammonium hydrazide and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry[J]. Analytical Biochemistry,2008,379:45-59
[101]Kang P., Mechref Y., Kyselova Z., et al. Comparative Glycomic Mapping through Quantitative Permethylation and Stable-Isotope Labeling[J]. Analytical Chemistry,2007, 79:6064-6073
[102]Weiskopf A.S., Vouros P., Harvey D.J. Characterization of oligosaccharide composition and structure by quadrupole ion trap mass spectrometry[J]. Rapid. Commun. Mass Spectrom.,1997,11:1493-1504
[103]Huntington J.A., Stein P.E. Structure and properties of ovalbumin[J]. Journal of Chromatography B,2001,756:189-198
[104]Harvey D. J. Postsource decay fragmentation of N-linked carbohydrates from ovalbumin and related glycoproteins[J]. American Society for Mass Spectrometry.2000, 11:572-577
[105]Harvey D. J., Wing D. R., Kuster B., et al. Composition of N-linked carbohydrates from ovalbumin and co-purified glycoproteins[J]. American Society for Mass Spectrometry, 2000,11:564-571
[106]廖秋林,陈晓东,丁彦青.血清蛋白质组学技术及研究进展[J].中国病理生理杂志,2008,24(2)：402-407
[107]Royle L., Campbell M. P., Radcliffe C. M., et al. Rapid and high-throughput analysis of N-glycans from ovarian cancer serum using a 96-well plate platform[J]. Analytical Biochemistry,2009,391:1-12
[108]周军,刘云叶,长明等.聚偏氟乙烯(PVDF)膜的制备及在水处理中应用的研究[J].新技术新产品,2007,(12)：51-55
[109]Masaaki Toyoda, Hiromi Ito, Yu-ki Matsuno, et al. Quantitative Derivatization of Sialic Acids for the Detection of Sialoglycans by MALDI MS[J]. Anal. Chem.,2008,80: 5211-5218
[110]Geoffrey R. Guile, Pauline M. Rudd, David R. Wing, et al. A Rapid High-Resolution High-Performance Liquid Chromatographic Method for Separating Glycan Mixtures and Analyzing Oligosaccharide Profiles[J], Analytical biochemistry,1996,240:210-226
[111]Lattova E., Varma S.,Bezabeh T., et al. Mass spectrometric profiling of N-Linked oligosaccharides and uncommon glycoform in mouse serum with head and neck tumor[J]. J Am Soc Mass Spectrom,2008,19:671-685
[112]Atwood J. A. Ⅲ, Cheng L., Alvarez-Manilla G., et al. Quantitation by isobaric labeling: applications to glycomics[J]. Journal of Proteome Research,2008, (7):367-374
[113]Harvey D. J., Royle L., Radcliffe C.M., et al. Structural and quantitative analysis of N-linked glycans by matrix-assisted laser desorption ionization and negative ion nanospray mass spectrometry[J], Analytical Biochemistry,2008,376:44-60
[114]Morten T. A., Simon M., Peter H. Site-Specific Glycoprofiling of N-Linked glycopeptides Using MALDI-TOF MS:strong correlation between signal strength and glycoform quantities[J]. Anal. Chem.,2009,81:3933-3943