Doss Jereme, Olubi Omotunde, Sannigrahi Biswajit, Williams M D, Gadi Deepti, Baird Barbara, Khan Ishrat
Department of Chemistry, Clark Atlanta University, Atlanta, GA, USA.
J Vis Exp. 2012 Sep 10(67):4135. doi: 10.3791/4135.
Electrospinning is an effective processing method for preparing nanofibers decorated with functional groups. Nanofibers decorated with functional groups may be utilized to study material-biomarker interactions i.e. act as biosensors with potential as single molecule detectors. We have developed an effective approach for preparing functional polymers where the functionality has the capacity of specifically binding with a model protein. In our model system, the functional group is 2,4-dinitrophenyl (DNP) and the protein is anti-DNP IgE (Immunoglobulin E). The functional polymer, α,ω-bi[2,4-dinitrophenyl caproic][poly(ethylene oxide)-b-poly(2-methoxystyrene)-b-poly(ethylene oxide)] (CDNP-PEO-P2MS-PEO-CDNP), is prepared by anionic living polymerization. The difunctional initiator utilized in the polymerization was prepared by electron transfer reaction of α-methylstyrene and potassium (mirror) metal. The 2-methoxystyrene monomer was added first to the initiator, followed by the addition of the second monomer, ethylene oxide, and finally the living polymer was terminated by methanol. The α,ω-dihydroxyl polymer [HO-PEO-P2MS-PEO-OH] was reacted with N-2,4-DNP-∈-amino caproic acid, by DCC coupling, resulting in the formation of α,ω-bi[2,4-dinitrophenylcaproic][poly(ethyleneoxide)-b-poly(2-methoxystyrene)-b-poly(ethylene oxide)] (CDNP-PEO-P2MS-PEO-CDNP). The polymers were characterized by FT-IR, (1)H NMR and Gel Permeation Chromatography (GPC). The molecular weight distributions of the polymers were narrow (1.1-1.2) and polymers with molecular weights greater than 50,000 was used in this study. The polymers were yellow powders and soluble in tetrahydrofuran. A water soluble CDNP-PEO-P2MS-PEO-CDNP/ DMEG (dimethoxyethylene glycol) complex binds and achieves steady state binding with solution IgE within a few seconds. Higher molecular weight (water insoluble i.e. around 50,000) CDNP-PEO-P2MS-PEO-CDNP polymers, containing 1% single wall carbon nanotubes (SWCNT) were processed into electroactive nanofibers (100 nm to 500 nm in diameter) on silicon substrate. Fluorescence spectroscopy shows that anti-DNP IgE interacts with the nanofibers by binding with the DNP functional groups decorating the fibers. These observations suggest that appropriately functionalized nanofibers hold promise for developing biomarker detection device.
静电纺丝是制备带有官能团的纳米纤维的一种有效加工方法。带有官能团的纳米纤维可用于研究材料与生物标志物的相互作用,即作为具有单分子探测器潜力的生物传感器。我们已经开发出一种制备功能聚合物的有效方法,其中的官能团能够与一种模型蛋白特异性结合。在我们的模型系统中,官能团是2,4 -二硝基苯基(DNP),蛋白是抗DNP IgE(免疫球蛋白E)。功能聚合物α,ω - 双[2,4 -二硝基苯基己酸][聚(环氧乙烷)-b -聚(2 -甲氧基苯乙烯)-b -聚(环氧乙烷)](CDNP - PEO - P2MS - PEO - CDNP)通过阴离子活性聚合制备。聚合中使用的双官能引发剂通过α -甲基苯乙烯与钾(镜面)金属的电子转移反应制备。首先将2 -甲氧基苯乙烯单体加入引发剂中,接着加入第二种单体环氧乙烷,最后用甲醇终止活性聚合物。α,ω - 二羟基聚合物[HO - PEO - P2MS - PEO - OH]通过DCC偶联与N - 2,4 - DNP - ε -氨基己酸反应,生成α,ω - 双[2,4 -二硝基苯基己酸][聚(环氧乙烷)-b -聚(2 -甲氧基苯乙烯)-b -聚(环氧乙烷)](CDNP - PEO - P2MS - PEO - CDNP)。这些聚合物通过傅里叶变换红外光谱(FT - IR)、核磁共振氢谱(¹H NMR)和凝胶渗透色谱(GPC)进行表征。聚合物的分子量分布较窄(1.1 - 1.2),本研究中使用的聚合物分子量大于50,000。这些聚合物为黄色粉末,可溶于四氢呋喃。一种水溶性的CDNP - PEO - P2MS - PEO - CDNP/二甲基乙二醇(DMEG)复合物在几秒钟内就能与溶液中的IgE结合并达到稳态结合。分子量较高(水不溶性,即约50,000)且含有1%单壁碳纳米管(SWCNT)的CDNP - PEO - P2MS - PEO - CDNP聚合物在硅基片上被加工成电活性纳米纤维(直径为100纳米至500纳米)。荧光光谱表明,抗DNP IgE通过与修饰在纤维上的DNP官能团结合而与纳米纤维相互作用。这些观察结果表明,适当功能化的纳米纤维在开发生物标志物检测设备方面具有前景。