Moody E D, Viskari P J, Colyer C L
Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109, USA.
J Chromatogr B Biomed Sci Appl. 1999 Jun 11;729(1-2):55-64. doi: 10.1016/s0378-4347(99)00121-8.
Indocyanine green (ICG) is a negatively charged, water-soluble, tricarbocyanine dye used primarily for medical imaging. ICG is only weakly fluorescent in the near-infrared region in its free (unbound) state in dilute aqueous solution. However, when non-covalently bound to protein, its fluorescence is greatly enhanced, making it a candidate for diode laser-induced fluorescence (diode-LIF) detection of proteins in capillary electrophoresis (CE). This paper investigates the suitability of ICG as a fluorescent label for the separation and detection of human serum albumin (HSA) by CE with diode-LIF detection. Specifically, we have considered the separation conditions necessary to resolve free ICG from ICG-HSA complexes; the limits of detection for free and HSA-bound ICG; the stability of aqueous ICG and ICG-HSA solutions over time; and the stoichiometry of the ICG-HSA complex.
吲哚菁绿（ICG）是一种带负电荷的水溶性三碳菁染料，主要用于医学成像。在稀水溶液中，处于游离（未结合）状态的ICG在近红外区域仅具有微弱的荧光。然而，当与蛋白质非共价结合时，其荧光会大大增强，这使其成为毛细管电泳（CE）中二极管激光诱导荧光（diode-LIF）检测蛋白质的候选物质。本文研究了ICG作为荧光标记物，用于通过CE结合二极管-LIF检测分离和检测人血清白蛋白（HSA）的适用性。具体而言，我们考虑了从ICG-HSA复合物中分离游离ICG所需的分离条件；游离ICG和与HSA结合的ICG的检测限；ICG水溶液和ICG-HSA溶液随时间的稳定性；以及ICG-HSA复合物的化学计量比。
