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在 SDS-CGE 中比较 LED 诱导荧光和激光诱导荧光:在抗体分析中的应用。

A comparative study of LED-induced fluorescence and laser-induced fluorescence in SDS-CGE: application to the analysis of antibodies.

机构信息

Picometrics, Toulouse, France.

出版信息

Electrophoresis. 2012 Jul;33(12):1709-14. doi: 10.1002/elps.201200132.

Abstract

LEDs present an alternative to lasers in LIF detection with CE, resulting in LED-induced fluorescence (LEDIF). LEDs are much less expensive, consume less energy and are more stable. In addition, LED light sources allow a greater range of wavelengths to better match the maximum wavelength for the fluorescence of the dye. Antibodies were largely studied in SDS capillary gel electrophoresis (SDS-CGE) and LIF detection with different dyes to label the proteins. In this work, our goal is to show that LEDs can advantageously replace lasers. We used 5-carboxytetramethylrhodamine succinimidyl ester (5-TAMRA.SE), 3-(2-furoyl)-quinoline-2 carboxaldehyde (FQ), and naphthalene-2,3-dialdehyde (NDA) to label IgG and we compared the LIF sensitivity with that obtained from LEDIF. We measured that the LOD values of LEDIF are identical to that obtained with the wavelength equivalent laser, and for 5-TAMRA.SE analysis, LOD values are about six times better than when the classical 488 nm laser was used.

摘要

LED 作为 CE 下 LIF 检测的激光替代方案,产生了 LED 诱导荧光(LEDIF)。LED 价格便宜得多,能耗低,稳定性也更高。此外,LED 光源允许更大的波长范围,以更好地匹配染料荧光的最大波长。抗体在 SDS 胶毛细管电泳(SDS-CGE)和不同染料的 LIF 检测中得到了广泛研究,用于标记蛋白质。在这项工作中,我们的目标是展示 LED 可以有利地替代激光。我们使用 5-羧基四甲基罗丹明琥珀酰亚胺酯(5-TAMRA.SE)、3-(2-呋喃基)-喹啉-2-甲醛(FQ)和萘-2,3-二醛(NDA)标记 IgG,并比较了 LIF 灵敏度与 LEDIF 的灵敏度。我们测量到 LEDIF 的 LOD 值与等效波长激光获得的 LOD 值相同,对于 5-TAMRA.SE 分析,LOD 值比使用经典的 488nm 激光时提高了约 6 倍。

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