5,5'-二硫代双(琥珀酰亚氨基-2-硝基苯甲酸)功能化的SERS纳米探针在免疫检验中的应用

doi:10.7503/cjcu20150116

高等学校化学学报 ›› 2015, Vol. 36 ›› Issue (8): 1505.doi: 10.7503/cjcu20150116

5,5'-二硫代双(琥珀酰亚氨基-2-硝基苯甲酸)功能化的SERS纳米探针在免疫检验中的应用

陈雷¹(), 薛向欣¹, 崔运成¹, 韩晓霞², 徐蔚青², 赵冰²

1. 吉林师范大学环境友好材料制备与应用教育部重点实验室, 四平 136000
2. 吉林大学超分子结构与材料国家重点实验室, 长春 130012

收稿日期:2015-02-02 出版日期:2015-08-10 发布日期:2015-07-17
作者简介:联系人简介: 陈雷, 男, 博士, 讲师, 主要从事基于SERS技术的蛋白质检测研究. E-mail:chenlei@jlnu.edu.cn
基金资助:
吉林省科技发展计划项目(批准号: 20150520015JH)、国家自然科学基金(批准号: 21403082, 21273091)和吉林师范大学博士科研启动项目(批准号: 2014004)资助

5,5'-Dithiobis(succinimidyl-2-nitrobenzoate) Functionalized Surface-enhanced Raman Scattering Nanoprobe for Immunoassay^†

CHEN Lei¹(), XUE Xiangxin¹, CUI Yuncheng¹, HAN Xiaoxia², XU Weiqing², ZHAO Bing²

1. Key Laboratory of Preparation and Applications of Environmental Friendly Materials,Ministry of Education,Jilin Normal University,Siping 136000, China
2. State Key Laboratory of Supramolecular Structure and Materials,Jilin University,Changchun 130012, China

Received:2015-02-02 Online:2015-08-10 Published:2015-07-17
Supported by:
† Supported by the Development Program of the Science and Technology of Jilin Province, China(No.20150520015JH), the National Natural Science Foundation of China(Nos.21403082, 21273091) and the Scientific Foundation for Young Scientists of Jilin Normal University, China(No.2014004)

摘要/Abstract

摘要：

以5,5'-二硫代双(琥珀酰亚氨基-2-硝基苯甲酸)(DSNB)分子作为偶联剂将蛋白质结合在金纳米粒子表面, 既保持了蛋白质的生物活性, 同时DSNB分子又具有较高的表面增强拉曼散射(SERS)活性, 可作为蛋白质定量分析的探针分子. 选用生物素与亲和素的特异性识别以及抗原抗体的免疫识别2个生物反应体系, 将SERS纳米探针固定在蛋白质检测芯片上. 以硅片为蛋白质检测载体, 利用硅片在520 cm^-1处的拉曼特征峰为内标, 对人IgG抗体进行定量分析. 结果表明, 该探针对人IgG抗体检测的最低浓度可以达到5 pg/mL.

关键词: 表面增强拉曼散射, 免疫检验, 纳米探针, 生物素, 链霉亲和素

Abstract:

A biofunctionalized nanoprobe for surface-enhanced Raman scattering(SERS)-based immunoassay was designed. 5,5'-Dithiobis(succinimidyl-2-nitrobenzoate)(DSNB) was employed as a linker for protein and gold nanoparticle, which can help to maintain the biological activity of the proteins. Besides that, DSNB possesses a high Raman scattering section, which can be used as the SERS probe as quantitative determination of proteins. Two kinds of biological systems(avidin-streptavidin and antigen-antibody) were selected for the target protein and nanoprobe specific binding to the protein chip. Meanwhile, silicon slide was employed as a carrier, which provides a Raman peak at 520 cm^-1 and can be used as a good internal standard for quantitative analysis of proteins. Using this method, anti-human IgG concentration in solutions can be quantitatively measured at concentrations as low as 5 pg/mL.

Key words: Surface-enhanced Raman scattering(SERS), Immunoassay, Nanoprobe, Avidin, Streptavidin

中图分类号:

O657

TrendMD:

陈雷, 薛向欣, 崔运成, 韩晓霞, 徐蔚青, 赵冰. 5,5'-二硫代双(琥珀酰亚氨基-2-硝基苯甲酸)功能化的SERS纳米探针在免疫检验中的应用. 高等学校化学学报, 2015, 36(8): 1505.

CHEN Lei, XUE Xiangxin, CUI Yuncheng, HAN Xiaoxia, XU Weiqing, ZHAO Bing. 5,5'-Dithiobis(succinimidyl-2-nitrobenzoate) Functionalized Surface-enhanced Raman Scattering Nanoprobe for Immunoassay^†. Chem. J. Chinese Universities, 2015, 36(8): 1505.

图/表 7

Scheme 1 Preparation of gold nanoprobe

Fig.1 Proposed biofunctional gold NPs for immunoassay

Fig.2 TEM image of gold NPs covered with DSNB and streptavidin(A) and SEM image of aggregated gold NPs after immunoassay(B)

Fig.3 UV-Vis spectra of DSNB(1×10-4 mol/L)(a), DSNB-streptavidin(100 μg/mL)(b), gold nano-particles covered with DSNB and streptavidin(c), gold nanoparticles(d) and gold nanoparticles covered with DSNB(e)

Fig.4 SERS spectra of DSNB before(a) and after(b—j) immunoassay with different concentrations of anti-human IgGConcentrations of anti-human IgG from b to j: 1×10-5, 5×10-6, 1×10-6, 5×10-7, 1×10-7, 5×10-8, 1×10-8, 5×10-9 and 0 mg/mL, respectively.

Fig.5 Concentration-dependent SERS intensities at 1336 cm-1 for DSNB containing different concentrations of anti-human IgG

Fig.6 Protein-induced SERS intensity changes of the DSNB-functionalized nanoprobe for immunoassay of anti-human IgG(5×10-7 mg/mL) detection systemFBS including 3×10-7—4×10-7 mg/mL protein; BSA: 5×10-7mg/mL; HSA: 5×10-7 mg/mL; Lys: 5×10-7 mg/mL.

参考文献 27

[1]	Jeanmaire D. L., van Duyne R. P., J. Electroanal. Chem., 1977, 84(1), 1—20
[2]	Albrecht M. G., Creighton J. A., J. Am. Chem. Soc., 1977, 99(15), 5215—5217
[3]	Aroca R., Surface-Enhanced Vibrational Spectroscopy, John Wiley & Sons, Chichester, 2006, 73—176
[4]	Kneipp K., Moskovits M., Kneipp H., Surface-Enhanced Raman Scattering-Physics and Applications, Springer, Berlin, Heidelberg, 2006, 335—349
[5]	Chen L., Han X. X., Yang J. X., Zhou J., Lü Z. C., Song W., Zhao B., Xu W. Q., Ozaki Y., Chem. Res. Chinese Universities, 2011, 27(4), 683—687
[6]	Moskovits M., Rev. Mod. Phys., 1985, 57(3), 783—826
[7]	Tian Z. Q., Ren B., Annu. Rev. Phys. Chem., 2004, 55(1), 197—229
[8]	Nie S. M., Emory S. R., Science, 1997, 275(5303), 1102—1106
[9]	Kneipp K., Wang Y., Kneipp H., Perelman L. T., Itzkan I., Dasari R. R., Feld M. S., Phys. Rev. Lett., 1997, 78(9), 1667—1670
[10]	Han X. X., Zhao B., Ozaki Y., Anal. Bioanal. Chem., 2009, 394(7), 1719—1727
[11]	Nathaniel L. R., Chad A. M., Chem. Rev., 2005, 105(4), 1547—1562
[12]	Katherine A. W., Anal. Bioanal. Chem., 2009, 394(1), 85—94
[13]	Han X. X., Huang G. G., Zhao B., Ozaki Y., Anal. Chem., 2009, 81(9), 3329—3333
[14]	Han X. X., Chen L., Ji W., Xie Y. F., Zhao B., Ozaki Y., Small, 2011, 7(3), 316—320
[15]	Jun B. H., Noh M. S., Kim J., Kim G., Kang H., Kim M. S., Seo Y. T., Baek J., Kim J. H., Park J., Kim S., Kim Y. K., Hyeon T., Cho M. H., Jeong D. H., Lee Y. S., Small, 2010, 6(1), 119—125
[16]	Wang G., Park H. Y., Lipert R. J., Porter M. D., Anal. Chem., 2009, 81(23), 9643—9650
[17]	Chen L., Han X. X., Yang J. X., Zhou J., Song W., Zhao B., Xu W. Q., Ozaki Y., J. Colloid Interface Sci., 2011, 360(2), 482—487
[18]	Han X. X., Kitahama Y., Tanaka Y., Guo J., Xu W. Q., Zhao B., Ozaki Y., Anal. Chem., 2008, 80(17), 6567—6572
[19]	Han X. X., Chen L., Guo J., Zhao B., Ozaki Y., Anal. Chem., 2010, 82(10), 4102—4106
[20]	Xu S. P., Ji W. Q., Xu W. Q., Li X. L., Wang L. Y., Bai Y. B., Zhao B., Ozaki Y., Analyst, 2004, 129(1), 63—68
[21]	Lin C. C., Yang Y. M., Chen Y. F., Yang T. S., Chang H. C., Biosens. Bioelectron., 2008, 24(2), 178—183
[22]	Chon H., Lee S., Son S. W., Oh C. H., Choo J. B., Anal. Chem., 2009, 81(8), 3029—3034
[23]	Wu Z. S., Zhou G. Z., Jiang J. H., Shen G. L., Yu R. Q., Talanta, 2006, 70(3), 533—539
[24]	Campbell F. M., Ingram A., Monaghan P., Cooper J., Sattar N., Eckersall P. D., Graham D., Analyst, 2008, 133(10), 1355—1357
[25]	Han X. X., Xie Y. F., Zhao B., Ozaki Y., Anal. Chem., 2010, 82(11), 4325—4328
[26]	Frens G., Nature Phys. Sci., 1973, 241(1), 20—22
[27]	Grubisha D. S., Lipert R. J., Park H. Y., Driskell J., Porter M. D., Anal. Chem., 2003, 75(21), 5936—5943

5,5'-二硫代双(琥珀酰亚氨基-2-硝基苯甲酸)功能化的SERS纳米探针在免疫检验中的应用

5,5'-Dithiobis(succinimidyl-2-nitrobenzoate) Functionalized Surface-enhanced Raman Scattering Nanoprobe for Immunoassay^†

RichHTML

PDF (PC)

可视化