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

doi:10.16183/j.cnki.jsjtu.2017.11.012

Abstract

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）

CLC Number:

TB 34

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.

References

［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.