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Carbon nanotube membranes for electrokinetic separation of proteins.

机译：碳纳米管膜，用于电动分离蛋白质。

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The purpose of this dissertation is to demonstrate the feasibility of using carbon nanotube membranes (CNTMs) to electrophoretically separate proteins based on their molecular weights (MW). The microfluidic device incorporating the CNTM was fabricated by traditional microfabrication techniques and thermal chemical vapor deposition (CVD). Preliminary electrophoretic testing indicated a lack of migration of fluorescent dye through a 1-mm long pad of CNTM due to the electrically conductive nature of CNTs. Efforts to minimize the conductive nature of the CNTs on the electric field prompted the use of 40-um long strips of CNTMs, which produced a voltage gradient similar to that of an empty channel and allowed complete migration of the dye. The fluorescently labeled non-denatured proteins of various MWs (6.5--- 97 kDa) were electrokinetically flowed through the microchannels in separate identical electriophoresis runs as well as in mixture form and observed through a fluorescent microscope. Proteins larger than or equal to 20 kDa accumulated ahead of the CNTMs, whereas, proteins smaller than or equal to 17 kDa freely migrated the CNTMs and no accumulation was observed, indicating that the protein build-up was due to the larger size of the higher MW proteins. More interestingly in addition to accumulation ahead of the CNTMs, mixtures of proteins larger than 20 kDa separated from each other and formed spatially-distinguished bands that were arranged in the increasing order of their MWs, such that the smallest protein accumulated against the CNTM, the next higher MW protein accumulated against the smallest protein and so on. Separation of protein components from 2-protein and 3-proteins mixtures (composed of Protein A [42 kDa], Neutravidin [66 kDa] and Phosphorylalse B [97 kDa], each labeled with a different fluorophore) was also verified by mapping pixel intensity of individual proteins to their locations inside the microchannel. The effect of CNTM geometry was evaluated and a 60 lm long CNTM strip in a 500 tm wide channel produced the best separation in terms of spatial discrimination between proteins.
机译：本文的目的是证明使用碳纳米管膜（CNTMs）基于分子量（MW）进行电泳分离的可行性。包含CNTM的微流控设备是通过传统的微制造技术和热化学气相沉积（CVD）来制造的。初步的电泳测试表明，由于CNT的导电特性，荧光染料没有通过1毫米长的CNTM垫迁移。尽量减少CNT在电场中的导电性的努力促使使用40um长的CNTM条，其产生的电压梯度类似于空通道的电压梯度，并允许染料完全迁移。各种分子量（6.5 --- 97 kDa）的荧光标记的非变性蛋白以电动方式流过微通道，并在单独的相同电泳电泳过程中以及以混合物形式流过微通道，并通过荧光显微镜观察。大于或等于20 kDa的蛋白质在CNTM之前积累，而小于或等于17 kDa的蛋白质则自由地迁移CNTMs，没有观察到积累，这表明蛋白质的积累是由于较高的分子大小所致。 MW蛋白。更有趣的是，除了在CNTM之前积累外，大于20 kDa的蛋白质混合物彼此分离并形成空间可区分的条带，并以其MW的增加顺序排列，从而使最小的蛋白质在CNTM上积累，下一个较高的MW蛋白相对最小的蛋白积累，依此类推。还通过绘制像素强度图验证了从2种蛋白质和3种蛋白质混合物（由A蛋白[42 kDa]，Neutravidin [66 kDa]和Phosphorylalse B [97 kDa]组成，分别用不同的荧光团标记）中分离蛋白质成分的方法。单个蛋白质在微通道内的位置。评估了CNTM几何形状的影响，就蛋白质之间的空间区分而言，在500 tm宽的通道中长60 lm的CNTM条带产生了最佳分离效果。

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