Creating biocompatible oil-water interfaces without synthesis: direct interactions between primary amines and carboxylated perfluorocarbon surfactants.
Creating biocompatible oil-water interfaces without synthesis: direct interactions between primary amines and carboxylated perfluorocarbon surfactants.
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DOI：
10.1021/ac4026048
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发表时间：
2013-11-05
影响因子：
7.4
通讯作者：
Easley, Christopher J.
中科院分区：
文献类型：
--
作者：
DeJournette, Cheryl J.;Kim, Joonyul;Medlen, Haley;Li, Xiangpeng;Vincent, Luke J.;Easley, Christopher J.
目前，用于赋予油包水液滴生物相容性的最突出的方法之一是合成由全氟聚醚和聚醚嵌段组成的三嵌段共聚物表面活性剂。将所得表面活性剂（EA表面活性剂、KryJeffa等）允许产生高度生物相容的液滴表面，同时保持起始材料的热稳定性。然而，这些表面活性剂的生产需要合成有机化学方面的专业知识，这对该领域的广泛采用造成了障碍。在这里，我们描述了一个简单的替代表面活性剂的合成改性，赋予生物相容性。我们已经观察到，仅通过利用水相中的聚醚二胺添加剂与油相中的羧化全氟化碳表面活性剂之间的结合相互作用，就可以使油包水液滴表面具有生物相容性和热稳定性。在这些条件下形成的液滴显示具有能够支持皮升规模的蛋白质测定、液滴聚合酶链式反应（PCR）和用等温重组酶聚合酶扩增（RPA）的液滴DNA扩增的生物相容性表面。用聚醚二胺水性添加剂形成的液滴足够稳定以承受PCR期间的温度循环（在60-94 °C下30-40个循环），同时保持生物相容性，并且RPA的反应效率显示与共价修饰的表面活性剂（KryJeffa）的反应效率相似。通过FT-IR光谱、NMR光谱、ESI-MS和荧光显微镜等多种方法证实了结合相互作用。总的来说，我们的研究结果表明，通过简单地将市售的聚醚二胺添加剂（Jeffamine艾德-900）引入水相，研究人员可以避免合成方法来生成能够支持亚纳升规模的DNA和蛋白质分析的生物相容性液滴表面。
Currently, one of the most prominent methods used to impart biocompatibility to aqueous-in-oil droplets is to synthesize a triblock copolymer surfactant composed of perfluoropolyether and polyether blocks. The resulting surfactants (EA surfactant, KryJeffa, etc.) allow generation of highly biocompatible droplet surfaces while maintaining the heat stability of the starting material. However, production of these surfactants requires expertise in synthetic organic chemistry, creating a barrier to widespread adoption in the field. Herein we describe a simple alternative to synthetic modification of surfactants to impart biocompatibility. We have observed that aqueous-in-oil droplet surfaces can be made biocompatible and heat stable by merely exploiting binding interactions between polyetherdiamine additives in the aqueous phase and carboxylated perfluorocarbon surfactants in the oil phase. Droplets formed under these conditions are shown to possess biocompatible surfaces capable of supporting picoliter-scale protein assays, droplet polymerase chain reaction (PCR), and droplet DNA amplification with isothermal recombinase polymerase amplification (RPA). Droplets formed with polyetherdiamine aqueous additives are stable enough to withstand temperature cycling during PCR (30–40 cycles at 60–94 °C) while maintaining biocompatibility, and the reaction efficiency of RPA is shown to be similar to that with a covalently-modified surfactant (KryJeffa). The binding interaction was confirmed with various methods, including FT-IR spectroscopy, NMR spectroscopy, ESI-MS, and fluorescence microscopy. Overall, our results suggest that by simply introducing a commercially–available, polyetherdiamine additive (Jeffamine ED-900) to the aqueous phase, researchers can avoid synthetic methods in generating biocompatible droplet surfaces capable of supporting DNA and protein analysis at the sub-nanoliter scale.
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