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Magnetized Carbon Nanotube Based Lateral Flow Immunoassay for Visual Detection of Complement Factor B

机译:基于磁化碳纳米管的横向流免疫测定用于目测补体因子B

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摘要

The authors describe a magnetized carbon nanotube (MCNT) based lateral flow immunoassay (LFI) for visual detection of complement factor B (CFB) in blood. MCNT was prepared by decorating magnetic Fe3O4 nanoparticles on multi-walled CNT surface and used as a colored tag for LFI. Monoclonal antibody (mAb, Ab1) of CFB was covalently immobilized on the MCNT surface via diimide-activated conjugation between the carboxyl groups on the MCNT surface and amino groups of antibodies. Polyclonal antibody of CFB (Ab2) and the secondary antibody were used to prepare the lateral flow test strips. The assay involved: (1) the capture of CFB in blood with the mAb-functionalized MCNT; (2) magnetic separation of the formed CFB-mAb-MCNT and excess of mAb-MCNT from the blood with an external magnet; (3) lateral flow test to capture the CFB-mAb-MCNT complex on the test zone and the excess of mAb-MCNT on the control zone; (4) Recording the intensities of the produced the characteristic brown bands with a portable strip reader and quantitating the concentration of CFB. The proof-of-concept was demonstrated by testing CFB in the buffer, and the detection limit was 5 ng mL−1 under the optimized analytical parameters. CFB in 1 μL of human blood was detected successfully in 30 min with this LFI and the results had a high correlation with commercial ELISA kit. Thence, the MCNT-based LFI offers a rapid and low-cost tool for detecting CFB in human blood directly.
机译:作者描述了一种基于磁化碳纳米管(MCNT)的侧向流动免疫分析(LFI),用于视觉检测血液中的补体因子B(CFB)。通过在多壁CNT表面上装饰磁性Fe3O4纳米颗粒来制备MCNT,并将其用作LFI的有色标签。 CFB的单克隆抗体(mAb,Ab1)通过MCNT表面的羧基与抗体的氨基之间的二酰亚胺活化共价共价固定在MCNT表面。 CFB(Ab2)多克隆抗体和二抗被用于制备侧流试纸条。该测定涉及:(1)用mAb功能化的MCNT捕获血液中的CFB; (2)利用外部磁体从血液中磁性分离形成的CFB-mAb-MCNT和过量的mAb-MCNT; (3)侧向流动测试,以捕获测试区的CFB-mAb-MCNT复合物和对照区的过量mAb-MCNT。 (4)用便携式条带读取器记录产生的特征性棕色带的强度,并定量CFB的浓度。通过在缓冲液中测试CFB证明了概念验证,在优化的分析参数下,检出限为5 ng mL -1 。使用此LFI在30分钟内成功检测到1μL人体血液中的CFB,其结果与市售ELISA试剂盒高度相关。因此,基于MCNT的LFI提供了一种快速,低成本的工具,可直接检测人血中的CFB。

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