会员登陆: 用户名:  密码: 验证码:
首页 杂志概况 投稿须知 在线投稿 在线阅读 征订启事 广告服务 行业资讯 企业动态 资料中心  专访报道 会展信息 ENGLISH

引用本文:   曾琼兰, 吴利, 李水明, 王勇. 纳升液相色谱-高分辨串联质谱研究不同分离方法对唾液外泌体蛋白质组的影响. 分析化学, 2018, 46(9): 1408-1414. doi:  10.11895/j.issn.0253-3820.181261 [复制]

Citation:   ZENG Qiong-Lan, WU Li, LI Shui-Ming, WANG Yong. Comparison of Separation Methods for Isolation in Analysis of Human Saliva-derived Exosomes Proteomics by Nano-liquid Chromatography-Mass Spectrometry. Chinese Journal of Analytical Chemistry, 2018, 46(9): 1408-1414. doi: 10.11895/j.issn.0253-3820.181261 [复制]

纳升液相色谱-高分辨串联质谱研究不同分离方法对唾液外泌体蛋白质组的影响

通讯作者:  王勇, wyong@szu.edu.cn

收稿日期: 2018-04-19

接受日期: 2018-07-19

出版日期: 2018-09-01

基金项目: 本文系深圳市科技计划项目(No.JCYJ20170818142154551)资助

Comparison of Separation Methods for Isolation in Analysis of Human Saliva-derived Exosomes Proteomics by Nano-liquid Chromatography-Mass Spectrometry

Corresponding author:  WANG Yong, wyong@szu.edu.cn

Received Date:  2018-04-19

Accepted Date:  2018-07-19

Published Date:  2018-09-01

Fund Project:  This work was supported by the Scientific Plan Program of Shenzhen, China (No. JCYJ20170818142154551)

外泌体是一系列胞外囊泡,是生物标志物的来源,但目前尚无灵敏高效的唾液外泌体蛋白质分离方法。本研究采用质谱方法比较唾液外泌体试剂盒(ExoQuick,EQ)方法和超高速离心(Ultracentrifugation,UC)方法分离外泌体的效果以及尿素缓冲液、RIPA裂解液、SDS裂解液提取外泌体蛋白质的效果。Brodford法和BCA定量测定结果表明,EQ方法分离0.5 mL唾液外泌体得到的蛋白质含量高于UC方法分离2 mL唾液所得到的蛋白质。进一步的质谱分析表明,前者鉴定到的蛋白质数目亦多于后者;试剂盒分离唾液外泌体与尿素缓冲液提取外泌体蛋白质的方法联用效果最佳,鉴定到194种蛋白质。本方法重现性好、样品用量少、检测结果稳定,可为在唾液外泌体中筛选疾病的标志物提供方法学参考。

关键词:   纳升液相色谱-高分辨串联质谱, 唾液外泌体, 外泌体试剂盒, 超高速离心法
Key words:   Nano liquid chromatography-high resolution tandem mass spectrometry, Saliva-exosome, ExoQuick, Ultracentrifugation
[1]

Thery C, Amigorena S, Raposo G, Clayton A. Curr. Protoc. Cell Biol., 2006, Chapter 3(Unit 3): 22-

[2]

Bobrie A, Colombo M, Raposo G, Thery C. Traffic, 2011, 12(12): 1659-1668. doi: 10.1111/j.1600-0854.2011.01225.x

[3]

Raposo G, Stoorvogel W. J. Cell Biol., 2013, 200(4): 373-383. doi: 10.1083/jcb.201211138

[4]

Azmi A S, Bao B, Sarkar F H. Cancer Metast. Rev., 2013, 32(3-4): 623-642. doi: 10.1007/s10555-013-9441-9

[5]

Li Y, St John M A, Zhou X, Kim Y, Sinha U, Jordan R C, Eisele D, Abemayor E, Elashoff D, Park N H, Wong D T. Clin. Cancer Res., 2004, 10(24): 8442-8450. doi: 10.1158/1078-0432.CCR-04-1167

[6]

Properzi F, Logozzi M, Fais S. Biomark Med., 2013, 7(5): 769-778. doi: 10.2217/bmm.13.63

[7]

Rabinowits G, Gerçeltaylor C, Day J M, Taylor D D, Kloecker G H. Clin. Lung Cancer, 2009, 10(1): 42-46. doi: 10.3816/CLC.2009.n.006

[8]

Shayne A, Belinda B, Bradley M, Andrew F. Front. Physiol., 2012, 3: 124-

[9]

Street J, Barran P E, Mackay C L, Weidt S, Balmforth C, Walsh T S, Chalmers R T, Webb D J, Dear J W. J. Transl. Med., 2012, 10: 5-. doi: 10.1186/1479-5876-10-5

[10]

Gonzalez-Begne M, Lu B, Han X, Hagen F K, Hand A R, Melvin J E, Yates J R. J. Proteome Res., 2009, 8(3): 1304-1314. doi: 10.1021/pr800658c

[11]

Kazuya I, Tamiko M, Kanako S, Yasutomo Y, Kiyotaka S. J. Extracell Vesicles, 2016, 5: 10-

[12]

Ogawa Y, Taketomi Y, Murakami M, Tsujimoto M, Yanoshita R. Biol. Pharm. Bull., 2013, 36(1): 66-75

[13]

Ogawa Y, Miura Y, Harazono A, Kanai-Azuma M, Akimoto Y, Kawakami H, Yamaguchi T, Toda T, Endo T, Tsubuki M, Yanoshita R. Biol. Pharm. Bull., 2011, 34(1): 13-23. doi: 10.1248/bpb.34.13

[14]

Lau C, Kim Y, Chia D, Spielmann N, Eibl G, Elashoff D, Wei F, Lin Y L, Moro A, Grogan T, Chiang S, Feinstein E, Schafer C, Farrell J, Wong D T. J. Biol. Chem., 2013, 288(37): 26888-26897. doi: 10.1074/jbc.M113.452458

[15]

Zlotogorski-Hurvitz A, Dayan D, Chaushu G, Korvala J, Salo T, Sormunen R, Vered M. J. Histochem. Cytochem., 2015, 63(3): 181-189. doi: 10.1369/0022155414564219

[16]

Zlotogorski-Hurvitz A, Dayan D, Chaushu G, Salo T, Vered M. Eur. J. Cancer, 2014, 50(11): S195-S196

[17]

Zlotogorski-Hurvitz A, Dayan D, Chaushu G, Salo T, Vered M. J. Cancer Res. Clin. Oncol., 2016, 142(1): 101-110. doi: 10.1007/s00432-015-2005-3

[18]

XU Jin-Ling, HONG Xiao-Yu, LI Shui-Ming, WANG Yong. Chinese J. Anal.Chem., 2016, 44(12): 1887-1891

徐金玲, 洪晓愉, 李水明, 王勇. 分析化学, 2016, 44(12): 1887-1891

[19]

Wišniewski J R, Zougman A, Nagaraj N, Mann M. Nat. Methods, 2009, 6(5): 359-362. doi: 10.1038/nmeth.1322

[20]

Chatterton R T J, Vogelsong K M, Lu Y C, Ellman A B, Hudgens G A. Clin. Physiol., 1996, 16(4): 433-448. doi: 10.1111/j.1475-097X.1996.tb00731.x

[21]

Ngoka L C M. Proteome Sci., 2008, 6: 30-. doi: 10.1186/1477-5956-6-30

计量
  • PDF下载量(16)
  • 文章访问量(116)
  • HTML全文浏览量(7)

目录

纳升液相色谱-高分辨串联质谱研究不同分离方法对唾液外泌体蛋白质组的影响

曾琼兰, 吴利, 李水明, 王勇

Figures and Tables