免疫质谱法检测肝癌血清多肽标志物

引用本文: 傅海媛, 原剑, 周晓明, 孙云波, 黄亚娟, 侯利平, 宋纯艳, 杨保安, 甄蓓, 高蓉, 貌盼勇, 林利忠, 徐淑芳, 肖汉族, 魏开华. 免疫质谱法检测肝癌血清多肽标志物[J]. 分析化学, 2012, 40(12): 1803-1806. doi: 10.3724/SP.J.1096.2012.20331 shu

Citation: FU Hai-Yuan, YUAN Jian, ZHOU Xiao-Ming, SUN Yun-Bo, HUANG Ya-Juan, HOU Li-Ping, SONG Cun-Yan, YANG Bao-An, ZHEN Pei, GAO Rong, MAO Pan-Yong, LIN Li-Zhong, XU Shu-Fang, XIAO Han-Zu, WEI Kai-Hua. Determination of Hepatocellular Carcinoma Serum Polypeptide Based on Antibody Preconcentration Coupled with Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, 2012, 40(12): 1803-1806. doi: 10.3724/SP.J.1096.2012.20331 shu

免疫质谱法检测肝癌血清多肽标志物

傅海媛 ^1,2, ,
原剑 ^2,3, ,
周晓明 ^2,3, ,
孙云波 ^2,3, ,
黄亚娟 ^2,3, ,
侯利平 ^2,3, ,
宋纯艳 ^2,3, ,
杨保安 ^4, ,
甄蓓 ^{2,3,
,} ,
高蓉 ^5, ,
貌盼勇 ^5, ,
林利忠 ^6, ,
徐淑芳 ^6, ,
肖汉族 ^6, ,
魏开华 ^{2,3,
,}

通讯作者: 魏开华,甄蓓; 魏开华,甄蓓

基金项目:
本文系卫生部传染病重大专项(No.2008ZX10002-016) (No.2008ZX10002-016)

863项目(No.2012AA020205)资助 (No.2012AA020205)

摘要: 建立免疫亲和富集分离与质谱分析相结合的免疫质谱法用于血清多肽标志物pep5的检测。多克隆抗体与蛋白A琼脂糖颗粒偶联用于捕获pep5,基质辅助激光解析电离飞行时间质谱检测标志物。本方法的线性范围为1-48 nmol/L,检出限0.2 nmol/L;回收率为93.0%-103.1%,相对标准偏差(RSD)为6.4%-11.8%;反复冻融3次pep5,检测结果稳定;检测临床肝癌/正常血清样本灵敏度为78.0%、特异度为90%。该方法方便、快速、准确,适用于临床样本中低浓度标志物的检测研究,对于癌症的早期诊断具有重要意义。

关键词:

English

Determination of Hepatocellular Carcinoma Serum Polypeptide Based on Antibody Preconcentration Coupled with Mass Spectrometry

1.
¹ 301 Militry Hospital of China, Beijing 100853, China;

Corresponding author: 魏开华,甄蓓; 魏开华,甄蓓

Received Date: 20 March 2012
Fund Project:
本文系卫生部传染病重大专项(No.2008ZX10002-016)

863项目(No.2012AA020205)资助

Abstract: Polypeptide antibody pre-concentration method coupled with mass spectrometry detecting was established for the determination of serum polypeptide. Polypeptide was captured by agarose beads followed by binding of specific polyclonal antibodies from serum. After timed washing steps, the remaining bound polypeptides were eluted from the beads and detected by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). We optimized a protein A agarose bead-based platform amenable to high throughput peptide capture followed by mass spectrometry can achieve ion signal, with precision (RSD<12%) and accuracy (about 10% of relative error) sufficient for quantifying biomarkers in the physiologically relevant nmol/L range. The method for the detection of peptide 5 (pep5) in the clinical hepatocellular carcinoma serum had sensitivity of 78.0% and specificity of 90%. The method is appropriate for the detection of low concentration biomarkers in clinical samples, and it will be significant for earlier diagnosis of cancers.

Key words:

1. [1]
  1 Sparbier K, Wenzel T, Dihazi H, Blaschke S, Müller GA, Deelder A, Flad T, Kostrzewa M. Proteomics, 2009, 9(6): 1442-14501 Sparbier K, Wenzel T, Dihazi H, Blaschke S, Müller GA, Deelder A, Flad T, Kostrzewa M. Proteomics, 2009, 9(6): 1442-1450
2. [2]
  2 YUAN Jian, SUN Yun-Bo, WEI Kai-Hua. Chinese J. Anal. Chem., 2011, 39(8): 1129-11332 YUAN Jian, SUN Yun-Bo, WEI Kai-Hua. Chinese J. Anal. Chem., 2011, 39(8): 1129-1133
3. [3]
  原剑, 孙云波, 魏开华. 分析化学, 2011,39(8): 1129-1133原剑, 孙云波, 魏开华. 分析化学, 2011,39(8): 1129-1133
4. [4]
  3 Jemal A, Bray F, Center M M, Ferlay J, Ward E, Forman D A. CA: Cancer Journal for Clinicians, 2011, 61(12): 69-903 Jemal A, Bray F, Center M M, Ferlay J, Ward E, Forman D A. CA: Cancer Journal for Clinicians, 2011, 61(12): 69-90
5. [5]
  4 Carter J D, Labean T H. J. Nucleic Acids. 2011, 2011: 926595. Epub 2011 Oct 94 Carter J D, Labean T H. J. Nucleic Acids. 2011, 2011: 926595. Epub 2011 Oct 9
6. [6]
  5 Cui J, Zhang K, Huang Q, Yu Y, Peng X. Anal. Chim. Acta, 2011, 688(1): 84-895 Cui J, Zhang K, Huang Q, Yu Y, Peng X. Anal. Chim. Acta, 2011, 688(1): 84-89
7. [7]
  6 Whiteaker J R, Zhao L, Zhang H Y, Feng L C, Piening B D, Anderson L, Paulovich A G. Anal. Biochem., 2007, 362(1): 44-546 Whiteaker J R, Zhao L, Zhang H Y, Feng L C, Piening B D, Anderson L, Paulovich A G. Anal. Biochem., 2007, 362(1): 44-54
8. [8]
  7 Stefaniuk P, Cianciara J, Wiercinska-Drapalo A. World J. Gastroenterol., 2010, 16(4): 418-4247 Stefaniuk P, Cianciara J, Wiercinska-Drapalo A. World J. Gastroenterol., 2010, 16(4): 418-424
9. [9]
  8 Anderson N L, Jackson A, Smith D, Hardie D, Borchers C, Pearson T W. Mol. Cell Proteomics, 2009, 8(5): 995-10058 Anderson N L, Jackson A, Smith D, Hardie D, Borchers C, Pearson T W. Mol. Cell Proteomics, 2009, 8(5): 995-1005
10. [10]
  9 Ziganshin R, Arapidi G, Azarkin I, Zaryadieva E, Alexeev D, Govorun V, Ivanov V. J. Proteomics, 2011, 74(5): 595-6069 Ziganshin R, Arapidi G, Azarkin I, Zaryadieva E, Alexeev D, Govorun V, Ivanov V. J. Proteomics, 2011, 74(5): 595-606
11. [11]
  10 Villanueva J, Shaffer D R, Philip J, Chaparro C A, Erdjument-Bromage H, Olshen A B, Fleisher M, Lilja H, Brogi E, Boyd J, Sanchez-Carbayo M, Holland E C, Cordon-Cardo C, Scher H I, Tempst P. J. Clin. Invest., 2006, 116(1): 271-28410 Villanueva J, Shaffer D R, Philip J, Chaparro C A, Erdjument-Bromage H, Olshen A B, Fleisher M, Lilja H, Brogi E, Boyd J, Sanchez-Carbayo M, Holland E C, Cordon-Cardo C, Scher H I, Tempst P. J. Clin. Invest., 2006, 116(1): 271-284

扫一扫看文章

计量

PDF下载量: 0
文章访问数: 312
HTML全文浏览量: 16

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

免疫质谱法检测肝癌血清多肽标志物

通讯作者: 魏开华,甄蓓; 魏开华,甄蓓

English

Determination of Hepatocellular Carcinoma Serum Polypeptide Based on Antibody Preconcentration Coupled with Mass Spectrometry

Corresponding author: 魏开华,甄蓓; 魏开华,甄蓓

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

免疫质谱法检测肝癌血清多肽标志物

通讯作者: 魏开华,甄蓓; 魏开华,甄蓓

English

Determination of Hepatocellular Carcinoma Serum Polypeptide Based on Antibody Preconcentration Coupled with Mass Spectrometry

Corresponding author: 魏开华,甄蓓; 魏开华,甄蓓

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs