表面等离子共振技术测定生长抑素含量

高等学校化学学报 ›› 2010, Vol. 31 ›› Issue (7): 1327.

表面等离子共振技术测定生长抑素含量

高锦航¹, 刘瑞¹, 童欢², 李献¹, 强鸥¹ , 唐承薇^1,2

1. 四川大学华西医院生物治疗国家重点实验室,
2. 消化内科, 成都 610041

收稿日期:2009-12-14 出版日期:2010-07-10 发布日期:2010-07-10
通讯作者: 唐承薇, 女, 博士, 教授, 博士生导师, 主要从事胃肠多肽基础及临床研究. E-mail: cwtang@medmail.com.cn
基金资助:
国家自然科学基金重点项目(批准号: 30330270)资助.

Quantification of Somatostatin with a Surface Plasmon Resonance-based Biosensor

GAO Jin-Hang¹, LIU Rui¹, TONG Huan², LI Xian¹, QIANG Ou¹, TANG Cheng-Wei^1,2*

1. State Key Laboratory of Biotherapy,
2. Departement of Gastroenterology, West China Hospital, Sichuan University, Chengdu 610041, China

Received:2009-12-14 Online:2010-07-10 Published:2010-07-10
Contact: TANG Cheng-Wei. E-mail: cwtang@medmail.com.cn
Supported by:
国家自然科学基金重点项目(批准号: 30330270)资助.

摘要/Abstract

摘要：

应用表面等离子共振(SPR)技术建立了一种快速检测生长抑素(SST)含量的方法, 在pH=5.0, SST浓度为75 μg/mL的最佳偶联条件下, SST在CM5芯片上的偶联值为1231.7 Response unit(RU). 选择117.7 nmol/L作为固定的大分子量抗体浓度, 用抗原-抗体-抗原结合法检测SST纯品, 在20~2000 pg范围内, SST纯品含量和RU值之间可建立直线相关的标准曲线, R=-0.824, p<0.05, 变异系数CV=1.3%. 所建立的方法测定SST含量的范围为20~2000 pg, 重复性好且可随时检测, 有望用于SST的临床即时检测.

关键词: 生长抑素定量检测; 表面等离子共振; 抗原-抗体-抗原结合法

Abstract:

A quick technique of quantifying somatostatin(SST) with surface plasmon resonance(SPR) on the principle of antigen-antibody-antigen assay was established. The optimal coupling of CM5 sensor chip for SST was indicated as 1231.4 response unit(RU) at pH=5.0, and the SST concentration of 75 μg/mL. A negative linear relationship between SST content(20—2000 pg) and RU was obtained using the high molecular mass antibody for SST at 117.7 nmol/L, R=-0.824, p<0.05, coefficient of variation 1.3%. This method can be used to detect SST content at a range from 20 to 2000 pg with good repetition and real-time, which would be useful for clinical diagnosis and intensive care of critically ill patients.

Key words: Quantification of somatostatin; Surface plasmon resonance; Antigen-antibody-antigen binding assay

TrendMD:

高锦航, 刘瑞, 童欢, 李献, 强鸥 , 唐承薇. 表面等离子共振技术测定生长抑素含量. 高等学校化学学报, 2010, 31(7): 1327.

GAO Jin-Hang, LIU Rui, TONG Huan, LI Xian, QIANG Ou, TANG Cheng-Wei*. Quantification of Somatostatin with a Surface Plasmon Resonance-based Biosensor. Chem. J. Chinese Universities, 2010, 31(7): 1327.

参考文献

[1]Brazeau P., Vale W., Burgus R., Ling N., Butcher M., Rivier J., Guillemin R.. Science[J],1973, 179: 77—79
[2]van Op den bosch J., Adriaensen D., van Nassauw L., Timmermans J. P.. Regul. Pept.[J], 2009, 156: 1—8
[3]Dalm V. A. S. H., Hofland L. J., Lamberts S. W. J.. Mol. Cell Endocrinol.[J], 2008, 286: 262—277
[4]Volante M., Rosas R., Allìa E., Granata R., Baragli A., Muccioli G., Papotti M.. Mol. Cell Endocrinol.[J], 2008, 286: 219—229
[5]Burgos-Ramos E., Hervás-Aguilar A., Aguado-Llera D., Puebla-Jiménez L., Hernández-Pinto A. M., Barrios V., Arilla-Ferreiro E.. Mol. Cell Endocrinol.[J], 2008, 286: 104—111
[6]Arimura A., Groot K., Taminato T., Ertan A., Akdamar K., Ryan J., Pienta K., Vinik A. I.. Peptides[J], 1981, 2: 271—274
[7]Queiroz D. M., Mendes E. N., Rocha G. A., Moura S. B., Resende L. M., Barbosa A. J., Coelho L. G., Passos M. C., Castro L. P., Oliveira C. A., Lima G. F.. Scand. J. Gastroenterol[J], 1993, 28: 858—864
[8]Weckbecker G., Lewis I., Albert R., Schmid H. A., Hoyer D., Bruns C.. Nat. Rev. Drug Discov.[J], 2003, 2: 999—1017
[9]Wilson W. D.. Science[J], 2002, 295: 2103—2105
[10]CHEN Shou-Hui(陈受惠), NI Yong-Nian(倪永年), YANG Fan(杨帆), YANG Xiu-Rong(杨秀荣). Chem. J. Chinese Universities(高等学校化学学报)[J], 2008, 29(1): 60—63
[11]JIN Wei, MU Ying, ZOU Xiang-yu, JIN Qin-han. Chem. Res. Chinese Universities[J], 2008, 24(1): 24—28
[12]BIAtechnology Handbook[M], Uppsala: BIACORE AB, 1998: 7—72
[13]Cannon M. J., Papalia G. A., Navratilova I., Fisher R. J., Roberts L. R., Worthy K. M., Stephen A. G., Marchesini G. R., Collins E. J., Casper D., Qiu H., Satpaev D., Liparoto S. F., Rice D. A., Gorshkova I. I., Darling R. J., Bennett D. B., Sekar M., Hommema E., Liang A. M., Day E. S., Inman J., Karlicek S. M., Ullrich S. J., Hodges D., Chu T., Sullivan E., Simpson J., Rafique A., Luginbuhl B., Westin S. N., Bynum M., Cachia P., Li Y. J., Kao D., Neurauter A., Wong M., Swanson M., Myszka D. G.. Anal. Biochem.[J], 2004, 330: 98—113
[14]Casper D., Bukhtiyarova M., Springman E. B.. Anal. Biochem.[J], 2004, 325: 126—136
[15]Campbella C. T., Kim G.. Biomaterials[J], 2007, 28: 2380—2392
[16]Shankaran D. R., Gobi K. V. A., Miura N.. Sens. Actuator B: Chem.[J], 2007, 121: 158—177
[17]Usami M., Mitsunaga K., Ohno Y.. J. Steroid. Biaochem. Mol.[J], 2002, 81: 47—55
[18]Li Y., Kobayashi M., Furui K., Soh N., Nakano K., Imato T.. Anal. Chim. Acta[J], 2006, 576: 77—83
[19]Dupont A., Coy D. H., Alvarado-Urbina G., Cté J., Meyers C. A., McManus J., Barden N., De Léan A., Labrie F.. Clin. Endocrinol.(Oxf.)[J], 1979, 10(1): 47—54
[20]WU Hao(吴浩), DENG Yi-Yun(邓一芸), TANG Cheng-Wei(唐承薇), KANG Yan(康焰), HUANG Ming-Hui(黄明慧), HU Ling(胡玲). Chinese Critical Care Medicine(中国危重病急救医学)[J], 2009, 21(5): 307—310