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Bioconjugation of IgG Secondary Antibodies to Polymer Dots for Multicolor Subcellular Imaging
ACS Applied Nano Materials ( IF 5.3 ) Pub Date : 2020-01-16 , DOI: 10.1021/acsanm.9b02292
Dan Wang 1, 2 , Jie Liu 2 , Zhihe Liu 2 , Zhe Zhang 2 , Zezhou Sun 2 , Changfeng Wu 2 , Guofeng Wang 1
Affiliation  

Multiplexed subcellular imaging has attracted widespread attention in biology and medicine. Because the fluorescence brightness and stability of organic dyes and fluorescent proteins are not sufficient, the development of fluorescent nanoparticles is of broad interest. Semiconducting polymer dots (Pdots) exhibit prominent luminescence properties. However, the Pdot-IgG bioconjugates are largely unexplored in biological imaging. Here, we developed multicolor Pdots covalently conjugated with IgG secondary antibodies for specific labeling of different subcellular structures. By comparing the two functional polymers, we found that the functionalization by poly(styrene-co-maleic anhydride) (PSMA) could reliably produce Pdot-IgG conjugates for specific subcellular labeling, while those by a comb-like amphiphilic polymer show strong nonspecific binding in an intracellular environment. In addition, we minimize nonspecific adsorption of the Pdot bioconjugates by using bovine serum albumin and poly(ethylene glycol) as passivation agents, respectively. We finally demonstrate that the Pdot-PSMA-IgGs hold great potential in multiplexed visualization of the subcellular structure.

中文翻译:

IgG二级抗体与聚合物点的生物缀合,用于多色亚细胞成像

多重亚细胞成像已在生物学和医学领域引起广泛关注。由于有机染料和荧光蛋白的荧光亮度和稳定性不足,因此荧光纳米颗粒的开发受到广泛关注。半导体聚合物点(Pdot)表现出突出的发光特性。但是,Pdot-IgG生物共轭物在生物学成像中还没有被开发。在这里,我们开发了与IgG二抗共价缀合的多色Pdot,用于不同亚细胞结构的特异性标记。通过比较这两个功能性聚合物,我们发现,通过功能化聚(苯乙烯--顺丁烯二酸酐(PSMA)可以可靠地产生用于特定亚细胞标记的Pdot-IgG缀合物,而那些由梳状两亲聚合物构成的缀合物在细胞内环境中表现出很强的非特异性结合。此外,我们分别通过使用牛血清白蛋白和聚乙二醇作为钝化剂来最大程度地减少Pdot生物缀合物的非特异性吸附。我们最终证明,Pdot-PSMA-IgGs在亚细胞结构的多重可视化中具有巨大潜力。
更新日期:2020-01-16
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