当前位置：
X-MOL 学术
›
Bioconjugate Chem.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Dye Tool Box for a Fluorescence Enhancement Immunoassay
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2018-01-08 00:00:00 , DOI: 10.1021/acs.bioconjchem.7b00731 Ursula Eisold 1 , Frank Sellrie 2, 3 , Henry Memczak 4 , Anika Andersson 3 , Jörg A. Schenk 2, 3 , Michael U. Kumke 1
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2018-01-08 00:00:00 , DOI: 10.1021/acs.bioconjchem.7b00731 Ursula Eisold 1 , Frank Sellrie 2, 3 , Henry Memczak 4 , Anika Andersson 3 , Jörg A. Schenk 2, 3 , Michael U. Kumke 1
Affiliation
|
Immunochemical analytical methods are very successful in clinical diagnostics and are nowadays also emerging in the control of food as well as monitoring of environmental issues. Among the different immunoassays, luminescence based formats are characterized by their outstanding sensitivity making this format especially attractive for future applications. The need for multiparameter detection capabilities calls for a tool box of dye labels in order to transduce the biochemical reaction into an optically detectable signal. Here, in a multiparameter approach each analyte may be detected by a different dye with a unique emission color (covering the blue to red spectral range) or a unique luminescence decay kinetics. In the case of a competitive immunoassay format for each of the different dye labels an individual antibody would be needed. In the present paper a slightly modified approach is presented using a 7-aminocoumarin unit as the basic antigen against which highly specific antibodies were generated. Leaving the epitope region in the dyes unchanged but introducing a side group in positon 3 of the coumarin system allowed us to tune the optical properties of the coumarin dyes without the necessity of new antibody generation. Upon modification of the parent coumarin unit the full spectral range from blue to deep red was accessed. In the manuscript the photophysical characterization of the coumarin derivatives and their corresponding immunocomplexes with two highly specific antibodies is presented. The coumarin dyes and their immunocomplexes were characterized by steady-state and time-resolved absorption as well as emission spectroscopy. Moreover, fluorescence depolarization measurements were carried out to complement the data stressing the different binding modes of the two antibodies. The binding modes were evaluated using the photophysics of 7-aminocoumarins and how it was affected in the respective immunocomplexes, namely, the formation of the intramolecular charge transfer (ICT) as well as the twisted intramolecular charge transfer (TICT). In contrast to other antibody–dye pairs reported a distinct fluorescence enhancement upon formation of the antibody–dye complex up to a factor of 50 was found. Because of the easy emission color tuning by tailoring the coumarin substitution for the antigen binding in nonrelevant position 3 of the parent molecule, a dye tool box is on hand which can be used in the construction of competitive multiparameter fluorescence enhancement immunoassays (FenIA).
中文翻译：
荧光增强免疫分析染料工具箱
免疫化学分析方法在临床诊断中非常成功，如今在食品控制和环境问题监测中也正在兴起。在不同的免疫测定中，基于发光的形式的特征在于其出色的灵敏度，使得这种形式对未来的应用特别有吸引力。对多参数检测功能的需求要求使用染料标签的工具箱，以便将生化反应转化为可光学检测的信号。在这里，在多参数方法中，每种分析物可以由具有独特发射颜色（覆盖蓝色到红色光谱范围）或独特发光衰减动力学的不同染料检测。在针对每种不同染料标记的竞争性免疫测定形式的情况下，将需要单独的抗体。在本文中，提出了一种略微修改的方法，该方法使用7-氨基香豆素单元作为基本抗原，针对该基本抗原可生成高度特异性的抗体。保留染料中的表位区域不变，但在香豆素系统的3号正位置引入一个侧基，使我们能够调节香豆素染料的光学性质，而无需产生新的抗体。修改母体香豆素单元后，即可访问从蓝色到深红色的整个光谱范围。在手稿中，香豆素衍生物及其具有两种高度特异性抗体的相应免疫复合物的光物理特性得到了体现。香豆素染料及其免疫复合物的特征在于稳态和时间分辨吸收以及发射光谱。而且，进行了荧光去极化测量，以补充强调两种抗体不同结合方式的数据。使用7-氨基香豆素的光物理来评估结合模式，以及其如何在各自的免疫复合物中受到影响，即分子内电荷转移（ICT）以及扭曲的分子内电荷转移（TICT）的形成。与其他抗体-染料对相反，报告发现抗体-染料复合物形成后荧光增强明显，达到了50倍。由于通过针对母体分子非相关位置3上的抗原结合量定制香豆素替代物，可以轻松进行发射颜色调节，
更新日期：2018-01-08
中文翻译：
荧光增强免疫分析染料工具箱
免疫化学分析方法在临床诊断中非常成功，如今在食品控制和环境问题监测中也正在兴起。在不同的免疫测定中，基于发光的形式的特征在于其出色的灵敏度，使得这种形式对未来的应用特别有吸引力。对多参数检测功能的需求要求使用染料标签的工具箱，以便将生化反应转化为可光学检测的信号。在这里，在多参数方法中，每种分析物可以由具有独特发射颜色（覆盖蓝色到红色光谱范围）或独特发光衰减动力学的不同染料检测。在针对每种不同染料标记的竞争性免疫测定形式的情况下，将需要单独的抗体。在本文中，提出了一种略微修改的方法，该方法使用7-氨基香豆素单元作为基本抗原，针对该基本抗原可生成高度特异性的抗体。保留染料中的表位区域不变，但在香豆素系统的3号正位置引入一个侧基，使我们能够调节香豆素染料的光学性质，而无需产生新的抗体。修改母体香豆素单元后，即可访问从蓝色到深红色的整个光谱范围。在手稿中，香豆素衍生物及其具有两种高度特异性抗体的相应免疫复合物的光物理特性得到了体现。香豆素染料及其免疫复合物的特征在于稳态和时间分辨吸收以及发射光谱。而且，进行了荧光去极化测量，以补充强调两种抗体不同结合方式的数据。使用7-氨基香豆素的光物理来评估结合模式，以及其如何在各自的免疫复合物中受到影响，即分子内电荷转移（ICT）以及扭曲的分子内电荷转移（TICT）的形成。与其他抗体-染料对相反，报告发现抗体-染料复合物形成后荧光增强明显，达到了50倍。由于通过针对母体分子非相关位置3上的抗原结合量定制香豆素替代物，可以轻松进行发射颜色调节，
京公网安备 11010802027423号