New Fluorescent Labels with Tunable Hydrophilicity for the Rational Design of Bright Optical Probes for Molecular Imaging
New Fluorescent Labels with Tunable Hydrophilicity for the Rational Design of Bright Optical Probes for Molecular Imaging
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DOI：
10.1021/bc4000349
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发表时间：
2013-07-01
影响因子：
4.7
通讯作者：
Resch-Genger, Ute
中科院分区：
文献类型：
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作者：
Pauli, Jutta;Licha, Kai;Resch-Genger, Ute
近红外区域明亮光学探针和染料-生物分子缀合物的合理设计需要荧光标记，当与识别单元结合或与目标相互作用时，荧光标记能够保持高荧光量子产率。由于亲水性控制的染料聚集与同源 FRET 结合是染料-蛋白质缀合物中主要的荧光失活途径之一，因此需要荧光标记以最小的染料聚集实现更高的标记程度。为了更好地了解控制染料间相互作用的因素，我们系统地研究了新型 xS-IDCC 系列对称五次甲基的信号相关光谱特性、亲水性和聚集行为，这些对称五次甲基配备有两个、四个和六个磺酸基团，以及这些染料与针对癌症相关表皮生长因子 (EGF) 受体的 IgG 和抗体西妥昔单抗 (ctx) 的选定缀合物，并与金标准进行了比较Cy5.5。 6S-IDCC 在 PBS 等水性介质中显示出 190 000 M-1 cm(-1) 的摩尔吸收系数和 0.18 的荧光量子产率 (Phi(f))，并且几乎没有聚集，我们可以识别出与 Cy5.5 具有类似良好性能的荧光团。 6S-IDCC 和 Cy5.5 的生物共轭产生了高发射性靶向探针，染料与蛋白质 (DIP) 比率的可比 Phi(f) 值为 0.29
The rational design of bright optical probes and dye-biomolecule conjugates in the NIR-region requires fluorescent labels that retain their high fluorescence quantum yields when bound to a recognition unit or upon interaction with a target. Because hydrophilicity-controlled dye aggregation in conjunction with homo-FRET presents one of the major fluorescence deactivation pathways in dye-protein conjugates, fluorescent labels are required that enable higher labeling degrees with minimum dye aggregation. Aiming at a better understanding of the factors governing dye-dye interactions, we systematically studied the signal-relevant spectroscopic properties, hydrophilicity, and aggregation behavior of the novel xS-IDCC series of symmetric pentamethines equipped with two, four, and six sulfonic acid groups and selected conjugates of these dyes with IgG and the antibody cetuximab (ctx) directed against the cancer-related epidermal growth factor (EGF) receptor in comparison to the gold standard Cy5.5. With 6S-IDCC, which displays a molar absorption coefficient of 190 000 M-1 cm(-1) and a fluorescence quantum yield (Phi(f)) of 0.18 in aqueous media like PBS and nearly no aggregation, we could identify a fluorophore with a similarly good performance as Cy5.5. Bioconjugation of 6S-IDCC and Cy5.5 yielded highly emissive targeted probes with comparable Phi(f) values of 0.29 for a dye-to-protein (DIP) ratio