APP下载

基于砷和汞离子标记SiO2@Au纳米探针的超敏多元免疫分析方法研究

2014-07-10李天华等

分析化学 2014年6期
关键词:原子荧光探针分析方法

李天华等

摘 要 构建了一种基于

4 结 论

本研究以砷和汞标记的复合纳米粒子作为信号探针,基于夹心免疫反应机理,构建了一种以氢化物发生原子荧光法为检测手段的新型多元免疫分析方法(SMIAs)。本方法具有以下特点:(1)可实现两种标志物同时检测。(2)大部分血清基底物质(蛋白、脂肪、无机盐等)没有原子荧光信号,故而本方法适用于复杂体系中TMs的测定

References

1 LIANG HongWei, SUN YuFen. Chin. J. Clinical Medical, 2001, 8(5): 495-497

梁红卫, 孙玉汾. 中国临床医学, 2001, 8(5): 495-497

2 Ni X G, Bai X F, Mao Y L, Shao Y F, Wu J X, Shan Y, Zhao P. European Journal of Surgical Oncology (EJSO), 2005, 31(2): 164-169

3 Zheng G, Patolsky F, Cui Y, Wang W U, Lieber C M. Nature Biotechnology, 2005, 23(10): 12941301

4 WU ShaoXiong, XING Zhi, CHEN HongBing, WANG Juan, FENG Lu. Chinese J. Anal. Chem., 2009, 37(5): 711-714

吴少雄, 邢 志, 陈红兵, 王 娟, 冯 璐. 分析化学, 2009, 37(5): 711-714

5 Momplaisir G M, Lei T, Marshall W D. Anal. Chem., 1994, 66(20): 3533-3539

6 Mohan D, Pittman Jr C U. J. Hazardous Materials, 2007, 142(1): 1-53

7 Belyakova L A, Lyashenko D Y, Shvets A N. Russian J. Phys. Chem. A, 2010, 84(4): 656-660

8 Li J, Gao H, Chen Z, Wei X, Yang C F. Anal. Chim. Acta, 2010, 665(1): 98-104

9 SanchezRodas D, Corns W T, Chen B, Stockwell P B. J. Anal. At. Spectrom., 2010, 25(7): 933-946

10 LizMarzn L M, Giersig M, Mulvaney P. Langmuir., 1996, 12(18): 4329-4335

11 Du Y, Guo S, Qin H, Dong S, Wang E. Chem. Commun., 2012, 48(6): 799-801

12 Hu Y, Noelck S J, Drezek R A. ACS Nano, 2010, 4(3): 1521-1528

13 Yang L Q, Ren X L, Tang F Q, Zhang L. Biosens. Biolectron., 2009, 25(4): 889-895

14 Frens G. Nature Phys. Sci., 1973, 241: 20-22

15 Zhu Q, Chai Y, Yuan R, Zhuo Y. Anal. Chim. Acta, 2013, 800: 22-28

16 Ge L, Yan J, Song X, Yan M, Ge S, Yu J. Biomaterials, 2012, 33(4): 1024-1031

17 Fu Z, Liu H, Ju, H. Anal. Chem., 2006, 78(19): 6999-7005

18 Kadirvelu K, Thamaraiselvi K, Namasivayam C. Sep. Purif. Technol., 2001, 24: 497-505

An Ultrasensitive Simultaneous Immunoassay Based on

Arsenic and Mercury Ions Labeled SiO2@Au Nanoparticle Probes

LI TianHua1, GAN Ning1, WU DaZhen1, JIN HaiJuan1, CAO YuTing*1, JIANG QianLi2

1(School of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, China)

2(Division of Laboratory in Nanfang Hospital of Nanfang Medical University, Guangzhou 510515, China)

Abstract An ultrasensitive immunoassay was developed based on As3+ and Hg2+ labeled SiO2@Au nanoparticles signal tags and hydride generationatomic fluorescence spectrometry (HGAFS) for the detection of carcinoembryonic antigen(CEA) and carbohydrate antigen 199 (CA 199) respectively. Firstly, amino SiO2@Au NPs were synthesized for selective absorption of As3+ and Hg2+ ions respectively. Subsequently,the secondary antibody (Ab2) of CEA and CA 199 was respectively labeled on As3+ or Hg2+ SiO2@Au NPs to prepare the corresponding signal tags for CEA and CA 199. Based on the sandwich immunoassay scheme, the tags, two antigen and corresponding first antibodies were bioconjugated on the bottom of 96well plate at room temperature to form the immunocomplex. After it was dissolved in alkali solution, As3+ and Hg2+ ions were released in solution and detected by HGAFS, which concentration was proportional with logarithms of CEA and CA 199. The reaction conditions were optimized and the tags were characterized. This assay was based on determination of the concentration of As3+ and Hg2+ for quantization of the corresponding CEA and CA 199 antigen. The assay showed a wide linear range from 0.001 to 100.0 μg/L for CEA and 0.01-80 U/mL for CA 199, and a lower detection limit of 0.5 ng/L and 0.005 U/mL respectively. This proposed method was used in real serums samples, the results were consistence with that by ELISA. The immunoassay showed three orders of magnitude of sensitivity lower than that of ELISA, which provides a promising simultaneous immunoassay for the early diagnosis of cancer .

Keywords Simultaneous immunoassay; Tumor markers; Arsenic and mercury ions labeled SiO2@Au nanoprobes; Hydride generationatomic fluorescence

(Received 14 January 2014; accepted 23 March 2014)

This work was supported by the National Natural Science Foundation of China (No. 30901367/C0811) and the National Natural Science Foundation of Zhejiang Province, China (Nos. LY12C20004, 2012C23101, 2011C23126)

摘 要 构建了一种基于

4 结 论

本研究以砷和汞标记的复合纳米粒子作为信号探针,基于夹心免疫反应机理,构建了一种以氢化物发生原子荧光法为检测手段的新型多元免疫分析方法(SMIAs)。本方法具有以下特点:(1)可实现两种标志物同时检测。(2)大部分血清基底物质(蛋白、脂肪、无机盐等)没有原子荧光信号,故而本方法适用于复杂体系中TMs的测定

References

1 LIANG HongWei, SUN YuFen. Chin. J. Clinical Medical, 2001, 8(5): 495-497

梁红卫, 孙玉汾. 中国临床医学, 2001, 8(5): 495-497

2 Ni X G, Bai X F, Mao Y L, Shao Y F, Wu J X, Shan Y, Zhao P. European Journal of Surgical Oncology (EJSO), 2005, 31(2): 164-169

3 Zheng G, Patolsky F, Cui Y, Wang W U, Lieber C M. Nature Biotechnology, 2005, 23(10): 12941301

4 WU ShaoXiong, XING Zhi, CHEN HongBing, WANG Juan, FENG Lu. Chinese J. Anal. Chem., 2009, 37(5): 711-714

吴少雄, 邢 志, 陈红兵, 王 娟, 冯 璐. 分析化学, 2009, 37(5): 711-714

5 Momplaisir G M, Lei T, Marshall W D. Anal. Chem., 1994, 66(20): 3533-3539

6 Mohan D, Pittman Jr C U. J. Hazardous Materials, 2007, 142(1): 1-53

7 Belyakova L A, Lyashenko D Y, Shvets A N. Russian J. Phys. Chem. A, 2010, 84(4): 656-660

8 Li J, Gao H, Chen Z, Wei X, Yang C F. Anal. Chim. Acta, 2010, 665(1): 98-104

9 SanchezRodas D, Corns W T, Chen B, Stockwell P B. J. Anal. At. Spectrom., 2010, 25(7): 933-946

10 LizMarzn L M, Giersig M, Mulvaney P. Langmuir., 1996, 12(18): 4329-4335

11 Du Y, Guo S, Qin H, Dong S, Wang E. Chem. Commun., 2012, 48(6): 799-801

12 Hu Y, Noelck S J, Drezek R A. ACS Nano, 2010, 4(3): 1521-1528

13 Yang L Q, Ren X L, Tang F Q, Zhang L. Biosens. Biolectron., 2009, 25(4): 889-895

14 Frens G. Nature Phys. Sci., 1973, 241: 20-22

15 Zhu Q, Chai Y, Yuan R, Zhuo Y. Anal. Chim. Acta, 2013, 800: 22-28

16 Ge L, Yan J, Song X, Yan M, Ge S, Yu J. Biomaterials, 2012, 33(4): 1024-1031

17 Fu Z, Liu H, Ju, H. Anal. Chem., 2006, 78(19): 6999-7005

18 Kadirvelu K, Thamaraiselvi K, Namasivayam C. Sep. Purif. Technol., 2001, 24: 497-505

An Ultrasensitive Simultaneous Immunoassay Based on

Arsenic and Mercury Ions Labeled SiO2@Au Nanoparticle Probes

LI TianHua1, GAN Ning1, WU DaZhen1, JIN HaiJuan1, CAO YuTing*1, JIANG QianLi2

1(School of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, China)

2(Division of Laboratory in Nanfang Hospital of Nanfang Medical University, Guangzhou 510515, China)

Abstract An ultrasensitive immunoassay was developed based on As3+ and Hg2+ labeled SiO2@Au nanoparticles signal tags and hydride generationatomic fluorescence spectrometry (HGAFS) for the detection of carcinoembryonic antigen(CEA) and carbohydrate antigen 199 (CA 199) respectively. Firstly, amino SiO2@Au NPs were synthesized for selective absorption of As3+ and Hg2+ ions respectively. Subsequently,the secondary antibody (Ab2) of CEA and CA 199 was respectively labeled on As3+ or Hg2+ SiO2@Au NPs to prepare the corresponding signal tags for CEA and CA 199. Based on the sandwich immunoassay scheme, the tags, two antigen and corresponding first antibodies were bioconjugated on the bottom of 96well plate at room temperature to form the immunocomplex. After it was dissolved in alkali solution, As3+ and Hg2+ ions were released in solution and detected by HGAFS, which concentration was proportional with logarithms of CEA and CA 199. The reaction conditions were optimized and the tags were characterized. This assay was based on determination of the concentration of As3+ and Hg2+ for quantization of the corresponding CEA and CA 199 antigen. The assay showed a wide linear range from 0.001 to 100.0 μg/L for CEA and 0.01-80 U/mL for CA 199, and a lower detection limit of 0.5 ng/L and 0.005 U/mL respectively. This proposed method was used in real serums samples, the results were consistence with that by ELISA. The immunoassay showed three orders of magnitude of sensitivity lower than that of ELISA, which provides a promising simultaneous immunoassay for the early diagnosis of cancer .

Keywords Simultaneous immunoassay; Tumor markers; Arsenic and mercury ions labeled SiO2@Au nanoprobes; Hydride generationatomic fluorescence

(Received 14 January 2014; accepted 23 March 2014)

This work was supported by the National Natural Science Foundation of China (No. 30901367/C0811) and the National Natural Science Foundation of Zhejiang Province, China (Nos. LY12C20004, 2012C23101, 2011C23126)

摘 要 构建了一种基于

4 结 论

本研究以砷和汞标记的复合纳米粒子作为信号探针,基于夹心免疫反应机理,构建了一种以氢化物发生原子荧光法为检测手段的新型多元免疫分析方法(SMIAs)。本方法具有以下特点:(1)可实现两种标志物同时检测。(2)大部分血清基底物质(蛋白、脂肪、无机盐等)没有原子荧光信号,故而本方法适用于复杂体系中TMs的测定

References

1 LIANG HongWei, SUN YuFen. Chin. J. Clinical Medical, 2001, 8(5): 495-497

梁红卫, 孙玉汾. 中国临床医学, 2001, 8(5): 495-497

2 Ni X G, Bai X F, Mao Y L, Shao Y F, Wu J X, Shan Y, Zhao P. European Journal of Surgical Oncology (EJSO), 2005, 31(2): 164-169

3 Zheng G, Patolsky F, Cui Y, Wang W U, Lieber C M. Nature Biotechnology, 2005, 23(10): 12941301

4 WU ShaoXiong, XING Zhi, CHEN HongBing, WANG Juan, FENG Lu. Chinese J. Anal. Chem., 2009, 37(5): 711-714

吴少雄, 邢 志, 陈红兵, 王 娟, 冯 璐. 分析化学, 2009, 37(5): 711-714

5 Momplaisir G M, Lei T, Marshall W D. Anal. Chem., 1994, 66(20): 3533-3539

6 Mohan D, Pittman Jr C U. J. Hazardous Materials, 2007, 142(1): 1-53

7 Belyakova L A, Lyashenko D Y, Shvets A N. Russian J. Phys. Chem. A, 2010, 84(4): 656-660

8 Li J, Gao H, Chen Z, Wei X, Yang C F. Anal. Chim. Acta, 2010, 665(1): 98-104

9 SanchezRodas D, Corns W T, Chen B, Stockwell P B. J. Anal. At. Spectrom., 2010, 25(7): 933-946

10 LizMarzn L M, Giersig M, Mulvaney P. Langmuir., 1996, 12(18): 4329-4335

11 Du Y, Guo S, Qin H, Dong S, Wang E. Chem. Commun., 2012, 48(6): 799-801

12 Hu Y, Noelck S J, Drezek R A. ACS Nano, 2010, 4(3): 1521-1528

13 Yang L Q, Ren X L, Tang F Q, Zhang L. Biosens. Biolectron., 2009, 25(4): 889-895

14 Frens G. Nature Phys. Sci., 1973, 241: 20-22

15 Zhu Q, Chai Y, Yuan R, Zhuo Y. Anal. Chim. Acta, 2013, 800: 22-28

16 Ge L, Yan J, Song X, Yan M, Ge S, Yu J. Biomaterials, 2012, 33(4): 1024-1031

17 Fu Z, Liu H, Ju, H. Anal. Chem., 2006, 78(19): 6999-7005

18 Kadirvelu K, Thamaraiselvi K, Namasivayam C. Sep. Purif. Technol., 2001, 24: 497-505

An Ultrasensitive Simultaneous Immunoassay Based on

Arsenic and Mercury Ions Labeled SiO2@Au Nanoparticle Probes

LI TianHua1, GAN Ning1, WU DaZhen1, JIN HaiJuan1, CAO YuTing*1, JIANG QianLi2

1(School of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, China)

2(Division of Laboratory in Nanfang Hospital of Nanfang Medical University, Guangzhou 510515, China)

Abstract An ultrasensitive immunoassay was developed based on As3+ and Hg2+ labeled SiO2@Au nanoparticles signal tags and hydride generationatomic fluorescence spectrometry (HGAFS) for the detection of carcinoembryonic antigen(CEA) and carbohydrate antigen 199 (CA 199) respectively. Firstly, amino SiO2@Au NPs were synthesized for selective absorption of As3+ and Hg2+ ions respectively. Subsequently,the secondary antibody (Ab2) of CEA and CA 199 was respectively labeled on As3+ or Hg2+ SiO2@Au NPs to prepare the corresponding signal tags for CEA and CA 199. Based on the sandwich immunoassay scheme, the tags, two antigen and corresponding first antibodies were bioconjugated on the bottom of 96well plate at room temperature to form the immunocomplex. After it was dissolved in alkali solution, As3+ and Hg2+ ions were released in solution and detected by HGAFS, which concentration was proportional with logarithms of CEA and CA 199. The reaction conditions were optimized and the tags were characterized. This assay was based on determination of the concentration of As3+ and Hg2+ for quantization of the corresponding CEA and CA 199 antigen. The assay showed a wide linear range from 0.001 to 100.0 μg/L for CEA and 0.01-80 U/mL for CA 199, and a lower detection limit of 0.5 ng/L and 0.005 U/mL respectively. This proposed method was used in real serums samples, the results were consistence with that by ELISA. The immunoassay showed three orders of magnitude of sensitivity lower than that of ELISA, which provides a promising simultaneous immunoassay for the early diagnosis of cancer .

Keywords Simultaneous immunoassay; Tumor markers; Arsenic and mercury ions labeled SiO2@Au nanoprobes; Hydride generationatomic fluorescence

(Received 14 January 2014; accepted 23 March 2014)

This work was supported by the National Natural Science Foundation of China (No. 30901367/C0811) and the National Natural Science Foundation of Zhejiang Province, China (Nos. LY12C20004, 2012C23101, 2011C23126)

猜你喜欢

原子荧光探针分析方法
硫脲浓度及反应时间对氢化物发生-原子荧光法测砷影响
荧光探针在游离肼检测中的研究进展
高层建筑沉降监测数据处理中多元回归分析方法的应用研究
高层建筑沉降监测数据处理中多元回归分析方法的应用研究
高分辨质谱在氯化石蜡分析方法中的应用
液相色谱—氢化物发生原子荧光光谱法测定饲料中砷的形态
超声提取—三氯化钛还原一原子荧光光谱法对土壤样品中不同价态碲的测定
例谈一种追及问题的错解和常用分析方法
通过接触测试来提高探针痕迹的一致性
并行测试探针卡的移动规则选择