主客体结构微球的流式免疫荧光检测的方法

doi:10.16183/j.cnki.jsjtu.2017.11.012

上海交通大学学报（自然版） ›› 2017, Vol. 51 ›› Issue (11): 1361-1366.doi: 10.16183/j.cnki.jsjtu.2017.11.012

主客体结构微球的流式免疫荧光检测的方法

吴志民，徐宏，古宏晨

上海交通大学生物医学工程学院，上海 200030

出版日期:2017-11-30 发布日期:2017-11-30
基金资助:
：国家高技术研究发展计划(863)项目（2012AA020103），上海市经信委产学研课题项目（CXY-2013-56）

Research on Immunofluorescent Detection Method by Flow Cytometry Based on Bead-on-Bead Structured Particles

WU Zhimin,XU Hong,GU Hongchen

School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China

Online:2017-11-30 Published:2017-11-30

摘要/Abstract

摘要： 采用纳米氧化硅客体子球和聚苯乙烯磁性主体母球设计并制备了一种新型的主客体组装结构复合微球，将抗人绒毛膜促性腺激素（hCG）抗体化学固载于组装微球表面，在流式免疫荧光检测平台中研究其免疫检测性能，并与传统的酶联免疫（ELISA）检测方法进行了比较.结果表明，主客体组装结构微球在流式免疫荧光检测中具有优异的检测性能，在微球用量极低的情况下仍然具有非常好的检测线性和检测重复性，并且空白限较传统的ELISA方法降低了6/7.

关键词: 主客体组装结构微球, 生物载体, 流式免疫荧光检测, 酶联免疫检测

Abstract: A new type host-guest structured bead-on-bead (BOB) particle was designed and synthesized via covalent assembling guest nano silica particles onto host magnetic polymer particles. Using BOB particles as biocarrier, anti-human chorionic gonadotrophin (hCG) antibody was covalently immobilized on the surface of BOB particles. Then, the immunofluorescent detection capability in recognition hCG antigen on flow cytometry was studied and compared with traditional enzyme-linked immuno sorbent assay (ELISA). The result showed that BOB particles presented remarkable immunofluorescent detection capability on flow cytometry. BOB particles also showed excellent linear detection than traditional ELISA detection in extremely low particles amount. Further more, the limit of blank (LOB) of immunofluorescent detection on flow cytometry was nearly 1/7 to traditional ELISA detection.

Key words: host-guest structured particles, bio-carrier, immunoflourescent detection, enzyme-linked immuno sorbent assay （ELISA）

中图分类号:

TB 34

吴志民，徐宏，古宏晨. 主客体结构微球的流式免疫荧光检测的方法[J]. 上海交通大学学报（自然版）, 2017, 51(11): 1361-1366.

WU Zhimin,XU Hong,GU Hongchen. Research on Immunofluorescent Detection Method by Flow Cytometry Based on Bead-on-Bead Structured Particles [J]. Journal of Shanghai Jiaotong University, 2017, 51(11): 1361-1366.

参考文献

［1］DUNBAR S A. Applications of Luminex xMAP technology for rapid, high-throughput multiplexed nucleic acid detection［J］. Clinica Chimica Acta, 2006, 363(1/2): 71-82. ［2］JUN B H, KANG H, LEE Y S, et al. Fluorescence-based multiplex protein detection using optically encoded microbeads［J］. Molecules, 2012, 17(3): 2474-2490. ［3］COLOMBO M, CARREGALROMERO S, CASULA M F, et al. Biological applications of magnetic nanoparticles［J］. Chemical Society Reviews, 2012, 41(11): 4306-4334. ［4］SHOJAEI T R, TABATABAEI M, SHAWKY S, et al. A review on emerging diagnostic assay for viral detection: The case of avian influenza virus［J］. Molecular Biology Reports, 2015, 42(1): 187-199. ［5］ZHANG H, LIU L, FU X, et al. Microfluidic beads-based immunosensor for sensitive detection of cancer biomarker proteins using multienzyme-nanoparticle amplification and quantum dots labels［J］. Biosensors & Bioelectronics, 2013, 42(1): 23-30. ［6］ZHU Ying, XU Hong, GU Hongchen. Progress of optically encoded microspheres for multiplexed assays［J］. Jourmal Shanghai Jiao Tong University (Science), 2014, 19(5): 521-530. ［7］DAI X, XU H, ZHANG X, et al. Determination of the affinity constant of streptavidin-coupled magnetic particles and a biotinylated antibody for high performance of magnetic solid carrier in immunoassays［J］. Materials Science & Engineering: C, 2014, 34(1): 422-428. ［8］ZHU Y, XU H, CHEN K, et al. Encoding through the host-guest structure: Construction of multiplexed fluorescent beads［J］. Chemical Communications, 2014, 50(90): 14041-14044. ［9］WANG Y F, ZHANG X, XU P, et al. Improvement of immunoassay detection sensitivity by using well-defined raspberry-like magnetic microbeads as carriers ［J］. Journal of Shanghai Jiao Tong University (Science), 2014, 19(5): 538-543. ［10］KAUR K, FORREST J A. Influence of particle size on the binding activity of proteins adsorbed onto gold nanoparticles［J］. Langmuir, 2012, 28(5): 2736-2744. ［11］ROACH P, FARRAR D, PERRY C C. Surface tailoring for controlled protein adsorption: Effect of topography at the nanometer scale and chemistry［J］. Journal of the American Chemical Society, 2006, 128(12): 3939-3945. ［12］ZHAO B, COLLINSON M M. Well-defined hierarchical templates for multimodal porous material fabrication［J］. Chemistry of Materials, 2010, 22(14): 4312-4319. ［13］王叶菲.拓扑状氧化硅/聚合物磁性纳米复合载体的可控制备及免疫检测应用研究［D］.上海：上海交通大学生物医学工程学院, 2014. ［14］YAMAGUCHI K, ITO M, TANIGUCHI T, et al. Preparation of core-shell composite polymer particles by a novel heterocoagulation based on hydrophobic interaction［J］. Colloid and Polymer Science, 2004, 282(4): 366-372. ［15］SAM S, TOUAHIR L, ANDRESA J S, et al. Semiquantitative study of the EDC/NHS activation of acid terminal groups at modified porous silicon surfaces ［J］. Langmuir, 2010, 26(2): 809-814.

主客体结构微球的流式免疫荧光检测的方法

Research on Immunofluorescent Detection Method by Flow Cytometry Based on Bead-on-Bead Structured Particles

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

编辑推荐

Metrics

本文评价