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Temperature and magnetic dual responsive microparticles for DNA separation

机译：温度和磁性双重响应微粒用于DNA分离

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The use of solid support in DNA separation from biological mixtures for diagnostics offers great potential for developing versatile separating tools. Although different polymer materials have been developed and studied for DNA separation, the application of such non-magnetic particles for DNA separation has remained limited. In this work, we describe the adsorption and desorption behavior of DNA on the temperature sensitive poly(N-isopropylacrylamide-co-aminoethylmethacrylate) (PNIPAM-co-AEMH) coated magnetic particles. Submicron sized temperature responsive magnetic/PDVB/P (NIPAM/AEMH/MBA) hybrid particles were prepared by a precipitation polymerization of NIPAM in the presence of previously prepared divinylbenzene (DVB) cross-linked magnetic particles as seed. Wherein, N,N-methylenebis-acrylamide (MBA), aminoethylmethacrylate hydrochloride (AEMH) and 2,2'-azobis (2-methylpropionam-idine) dihydrochloride (V-50) were used as a crosslinker, functional monomer and initiator respectively. The amino (-NH2) functional groups derived from AEMH and V-50 are able to be cationic charged (-NH_3~+) by being protonated and can interact electrostatically with negatively charged DNA. The adsorption and desorption behavior of Herring Sperm DNA onto these functional magnetic particles were investigated as a function of pH, temperature, time and ionic strength. Interestingly, it was found that there is a significant effect of temperature on the DNA adsorption onto these magnetic particles. The results obtained indicate that the maximum amount of DNA adsorption was found at lower salt concentration, acidic pH condition and below the volume phase transition temperature (VPTT) of the PNIPAM shell. The highest desorption was found at higher salt concentration in basic pH and above the VPTT (40 °C) temperature. The temperature responsive magnetic particles and procedure described here can be used in microsystem separation technology for nucleic acid extraction, purification and concentration.
机译：固体支持物用于从生物混合物中分离DNA进行诊断的用途为开发多功能分离工具提供了巨大的潜力。尽管已经开发和研究了用于DNA分离的不同聚合物材料，但是这种非磁性颗粒用于DNA分离的应用仍然受到限制。在这项工作中，我们描述了DNA在温度敏感的聚（N-异丙基丙烯酰胺-共-氨基乙基甲基丙烯酸酯）（PNIPAM-co-AEMH）涂覆的磁性颗粒上的吸附和解吸行为。通过在先前制备的二乙烯基苯（DVB）交联磁性颗粒作为种子的存在下，通过NIPAM的沉淀聚合反应，制备了亚微米级的温度响应磁性/ PDVB / P（NIPAM / AEMH / MBA）杂化颗粒。其中，将N，N-亚甲基双丙烯酰胺（MBA），甲基丙烯酸氨基乙酯盐酸盐（AEMH）和2，2'-偶氮双（2-甲基丙酰胺-吡啶）二盐酸盐（V-50）用作交联剂，功能单体和引发剂。源自AEMH和V-50的氨基（-NH2）官能团能够通过质子化而带阳离子电荷（-NH_3〜+），并且可以与带负电荷的DNA静电相互作用。研究了鲱鱼精子DNA在这些功能性磁性粒子上的吸附和解吸行为，该行为是pH，温度，时间和离子强度的函数。有趣的是，发现温度对DNA吸附在这些磁性颗粒上的影响很大。获得的结果表明，在较低的盐浓度，酸性pH条件和低于PNIPAM壳的体积相变温度（VPTT）时，发现最大的DNA吸附量。在碱性pH值和高于VPTT（40°C）温度的较高盐浓度下，发现最高的脱附率。本文所述的温度响应性磁性颗粒和程序可用于微系统分离技术中，以进行核酸提取，纯化和浓缩。

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