Highly robust and optimized conjugation of antibodies to nanoparticles using quantitatively validated protocols
Highly robust and optimized conjugation of antibodies to nanoparticles using quantitatively validated protocols
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DOI：
10.1039/c6nr04683e
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发表时间：
2017-02-21
期刊：
影响因子：
6.7
通讯作者：
Jeong, Dae Hong
中科院分区：
文献类型：
--
作者：
Jeong, Sinyoung;Park, Ji Yong;Jeong, Dae Hong
抗体偶联纳米颗粒（Antibody-conjugated nanoparticles，NPs）由于其对生物靶点的高灵敏度和特异性以及广泛的适用性而在诊断和治疗应用中引起了极大的关注。不幸的是，这些特征在缀合效率、抗体中靶结合位点的取向和化学缀合反应期间的变性方面受到抗体缀合方法的显著影响。此外，每个NP上缀合的抗体的数量和总体靶向功效是用抗体缀合的NP进行定量生物测定的关键因素。在此，我们报告了一种通用的和定向的抗体偶联方法，使用无铜点击化学。此外，使用荧光标记的抗体和抗原定量地和实验性地评价缀合抗体的数目和它们的结合能力。使用基于无铜点击化学的缀合策略制备的抗体缀合的NP的强结合能力比使用EDC/NHS偶联方法后观察到的结合能力高8倍上级。此外，开发的抗体缀合方法的通用性也通过抗体与三种二氧化硅包封的NP的缀合来证明。
Antibody-conjugated nanoparticles (NPs) have attracted great attention in diagnostic and therapeutic applications due to their high sensitivity and specificity for biotargets, as well as their wide applicability. Unfortunately, these features are significantly affected by antibody conjugation methods in terms of conjugation efficiency, orientation of the target binding site in the antibody, and denaturation during chemical conjugation reactions. Furthermore, the number of conjugated antibodies on each NP and the overall targeting efficacy are critical factors for a quantitative bioassay with antibody-conjugated NPs. Herein, we report a versatile and oriented antibody conjugation method using copper-free click chemistry. Moreover, the number of conjugated antibodies and their binding capacity were quantitatively and experimentally evaluated using fluorescently-labeled antibodies and antigens. The strong binding capability of antibody-conjugated NPs prepared using the copper-free click chemistry-based conjugation strategy was 8 times superior to the binding capability seen following the use of the EDC/NHS-coupling method. Additionally, the versatility of the developed antibody conjugation method was also demonstrated by conjugation of the antibody to three kinds of silica-encapsulated NPs.