Chemistry Department, Faculty of Science, Ferdowsi University of Mashhad, Mashhad 9177948974, Iran.
Chemistry Department, Faculty of Science, Ferdowsi University of Mashhad, Mashhad 9177948974, Iran; Nano Research Center, Ferdowsi University of Mashhad, Mashhad 9177948974, Iran.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Jun 15;1118-1119:63-69. doi: 10.1016/j.jchromb.2019.04.030. Epub 2019 Apr 16.
Despite the fast development of novel and high-resolution electrophoresis techniques such as capillary-based methods and microfluidic devices, the slab gel electrophoresis is still a popular method for the separation of biomolecules in medicine and biology. It is a low cost and simple method and offers high throughput. However, this technique is limited to low voltages leading to slow separations. Producing the heat during the electrophoresis known as Joule heating inevitably leads to a rise in the gel temperature. For the first time, this work offers a whole gel temperature measurement by using a thermal camera which presents accurate temperature profiles in the gel with a resolution of more than 10 pixel/mm and a precision of 0.1 °C. Titania, TiO, nanoparticles (NPs) were embedded into the polyacrylamide (PA) gel to improve the electrophoretic separation of proteins. By embedding 0.025% w/v TiO NPs, heat dissipation increases by 16.5% at applied voltage of 200 V compared with that of PA gel with no embedded TiO NPs. The thermal images showed that the composite gel was 2.5 °C in average cooler than PA gel after 15 min of electrophoresis run at 200 V. The maximum separation voltage increased by 30 V in the composite PA/TiO gel compared with the pure PA gel. Moreover, the average number of theoretical plates over the 10 protein peaks, as a criterion of separation performance, increased by about 63% at 180 V when TiO NPs were included into the gel.
尽管新型和高分辨率电泳技术(如基于毛细管的方法和微流控设备)发展迅速,但平板凝胶电泳仍然是医学和生物学中生物分子分离的常用方法。它是一种低成本、简单的方法,具有高通量。然而,该技术仅限于低电压,导致分离速度较慢。电泳过程中产生的焦耳热不可避免地导致凝胶温度升高。这项工作首次使用热像仪进行整个凝胶温度测量,其分辨率超过 10 像素/毫米,精度为 0.1°C,提供了凝胶中的准确温度分布。将纳米二氧化钛(TiO)颗粒嵌入聚丙烯酰胺(PA)凝胶中,以改善蛋白质的电泳分离。与未嵌入 TiO NPs 的 PA 凝胶相比,在 200 V 的应用电压下,嵌入 0.025% w/v TiO NPs 可使散热增加 16.5%。热图像显示,在 200 V 下电泳 15 分钟后,复合凝胶的平均温度比 PA 凝胶低 2.5°C。与纯 PA 凝胶相比,在复合 PA/TiO 凝胶中,最大分离电压增加了 30 V。此外,当将 TiO NPs 纳入凝胶中时,在 180 V 下,作为分离性能标准的 10 个蛋白质峰的平均理论塔板数增加了约 63%。