A Novel Near-infrared Fluorescent Probe for Detection of Aβ40 Aggregates Enhanced Publication

{ "defaultlang": "zh", "titlegroup": { "articletitle": [ { "lang": "zh", "data": [ { "name": "text", "data": "一种新型近红外荧光探针用于Aβ40聚集体的检测" } ] }, { "lang": "en", "data": [ { "name": "text", "data": "A Novel Near-infrared Fluorescent Probe for Detection of Aβ40 Aggregates" } ] } ] }, "contribgroup": { "author": [ { "name": [ { "lang": "zh", "surname": "徐", "givenname": "蕴泽", "namestyle": "eastern", "prefix": "" }, { "lang": "en", "surname": "XU", "givenname": "Yun-ze", "namestyle": "eastern", "prefix": "" } ], "stringName": [], "aff": [ { "rid": "aff1", "text": "1" }, { "rid": "aff2", "text": "2" } ], "role": [ "first-author" ], "bio": [ { "lang": "zh", "text": [ "徐蕴泽（1997-），男，山东青岛人，硕士研究生，2019年于安徽农业大学获得学士学位，主要从事有机荧光探针分子的合成及在阿尔茨海默症检测中应用的研究。 E-mail： xuyunze@zjnu.edu.cn" ], "graphic": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471839&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471847&type=", "big": 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[], "bio": [ { "lang": "zh", "text": [ "吕光磊（1986-），男，山东泰安人，博士，讲师，2017年于复旦大学获得博士学位，主要从事有机无机杂化材料的设计合成以及在疾病诊疗中应用的研究。 E-mail： guanglei@zjnu.edu.cn" ], "graphic": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471851&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471861&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471856&type=", "width": "24.89200020", "height": "29.88732910", "fontsize": "" } ], "data": [ [ { "name": "text", "data": "吕光磊" }, { "name": "text", "data": "（1986-），男，山东泰安人，博士，讲师，2017年于复旦大学获得博士学位，主要从事有机无机杂化材料的设计合成以及在疾病诊疗中应用的研究。 E-mail： " }, { "name": "text", "data": "guanglei@zjnu.edu.cn" } ] ] } ], "email": "guanglei@zjnu.edu.cn", "deceased": false }, { "name": [ { "lang": "zh", "surname": "李", "givenname": "春霞", "namestyle": "eastern", "prefix": "" }, { "lang": "en", "surname": "LI", "givenname": "Chun-xia", "namestyle": "eastern", "prefix": "" } ], "stringName": [], "aff": [ { "rid": "aff1", "text": "1" } ], "role": [], "bio": [ { "lang": "zh", "text": [ "李春霞（1977-），女，山东德州人，博士，教授，博士生导师，2008年于中国科学院长春应用化学研究所获得博士学位，主要从事功能纳米材料在荧光检测和肿瘤诊疗中的研究。E‑mail: cxli@sdu.edu.cn" ], "graphic": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471866&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471877&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471870&type=", "width": "24.89199448", "height": "29.88732910", "fontsize": "" } ], "data": [ [ { "name": "text", "data": "李春霞" }, { "name": "text", "data": "（1977-），女，山东德州人，博士，教授，博士生导师，2008年于中国科学院长春应用化学研究所获得博士学位，主要从事功能纳米材料在荧光检测和肿瘤诊疗中的研究。E‑mail: cxli@sdu.edu.cn" } ] ] } ], "deceased": false } ], "aff": [ { "id": "aff1", "intro": [ { "lang": "zh", "label": "1", "text": "山东大学 前沿交叉科学青岛研究院， 分子科学与工程研究院， 山东 青岛　266237", "data": [ { "name": "text", "data": "山东大学 前沿交叉科学青岛研究院， 分子科学与工程研究院， 山东 青岛　266237" } ] }, { "lang": "en", "label": "1", "text": "Institute of Molecular Sciences Engineering， Institute of Frontier and Interdisciplinary Science， Shandong University， Qingdao 266237， China", "data": [ { "name": "text", "data": "Institute of Molecular Sciences Engineering， Institute of Frontier and Interdisciplinary Science， Shandong University， Qingdao 266237， China" } ] } ] }, { "id": "aff2", "intro": [ { "lang": "zh", "label": "2", "text": "浙江师范大学 化学与生命科学学院， 浙江 金华　321004", "data": [ { "name": "text", "data": "浙江师范大学 化学与生命科学学院， 浙江 金华　321004" } ] }, { "lang": "en", "label": "2", "text": "College of Chemistry and Life Science， Zhejiang Normal University， Jinhua 321004， China", "data": [ { "name": "text", "data": "College of Chemistry and Life Science， Zhejiang Normal University， Jinhua 321004， China" } ] } ] } ] }, "abstracts": [ { "lang": "zh", "data": [ { "name": "p", "data": [ { "name": "text", "data": "β⁃淀粉样蛋白（Aβ）形成的老年斑块是阿尔茨海默症（Alzheimer's disease，AD）最重要的病理性标志物之一，在AD的发病中起到了非常重要的作用。在AD的发病前期，Aβ40聚集体是最为主要且大量存在的蛋白形式。因此，对Aβ40聚集体的检测有利于实现对AD的早期诊断。本工作设计合成了一种具有D⁃π⁃A结构的小分子近红外荧光探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C" } ] }, { "name": "text", "data": "⁃" }, { "name": "bold", "data": [ { "name": "text", "data": "1" } ] }, { "name": "text", "data": "用于Aβ40聚集体的荧光检测。探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C" } ] }, { "name": "text", "data": "⁃" }, { "name": "bold", "data": [ { "name": "text", "data": "1" } ] }, { "name": "text", "data": "具有良好的荧光发射波长（" }, { "name": "italic", "data": [ { "name": "text", "data": "λ" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "em" } ] }, { "name": "text", "data": "=640 nm），对Aβ40聚集体表现出了较好的结合能力（" }, { "name": "italic", "data": [ { "name": "text", "data": "K" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "d" } ] }, { "name": "text", "data": "=3.072 μmol/L），能够实现在溶液中对Aβ40聚集体的特异性检测。除此之外，该探针合成简单，成本低廉，灵敏度较高，响应速度极快。" } ] } ] }, { "lang": "en", "data": [ { "name": "p", "data": [ { "name": "text", "data": "β-Amyloid（Aβ） senile plaques are one of the most important pathological markers of Alzheimer's disease（AD）， which plays a quite important role in the pathogenesis of AD. In the early stage of AD， Aβ40 aggregates are the most predominant and abundant. Therefore， the detection of Aβ40 aggregates is conducive to the early diagnosis of AD. In this work， a small molecule fluorescent probe " }, { "name": "bold", "data": [ { "name": "text", "data": "C" } ] }, { "name": "text", "data": "-" }, { "name": "bold", "data": [ { "name": "text", "data": "1" } ] }, { "name": "text", "data": " with a D-π-A structure was designed and synthesized for the fluorescence detection of Aβ40 aggregates. The probe " }, { "name": "bold", "data": [ { "name": "text", "data": "C" } ] }, { "name": "text", "data": "-" }, { "name": "bold", "data": [ { "name": "text", "data": "1" } ] }, { "name": "text", "data": " displays superior fluorescence properties（" }, { "name": "italic", "data": [ { "name": "text", "data": "λ" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "em" } ] }, { "name": "text", "data": "=640 nm） and shows remarkable binding capacity for Aβ40 aggregates（" }, { "name": "italic", "data": [ { "name": "text", "data": "K" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "d" } ] }, { "name": "text", "data": "=3.072 μmol/L）， enabling the specific detection of Aβ40 aggregates in solution. In addition， the probe has series of advantages， including simple synthesis， low cost， high sensitivity and fast response." } ] } ] } ], "keyword": [ { "lang": "zh", "data": [ [ { "name": "text", "data": "荧光探针" } ], [ { "name": "text", "data": "Aβ" } ], [ { "name": "text", "data": "阿尔茨海默症" } ], [ { "name": "text", "data": "近红外" } ] ] }, { "lang": "en", "data": [ [ { "name": "text", "data": "fluorescent probe" } ], [ { "name": "text", "data": "Aβ" } ], [ { "name": "text", "data": "Alzheimer's disease" } ], [ { "name": "text", "data": "near-infrared （NIR）" } ] ] } ], "highlights": [], "body": [ { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "1　引　　言" } ], "level": "1", "id": "s1" } }, { "name": "p", "data": [ { "name": "text", "data": "阿尔茨海默症（Alzheimer's disease，AD）是一种常见的神经退行性疾病，与癌症、心脑血管疾病一起成为当下危害人类健康的最主要的三大疾病" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "1", "type": "bibr", "rid": "R1", "data": [ { "name": "text", "data": "1" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。阿尔茨海默症主要发病于老年人群体，严重威胁着老年人的身心健康" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "2", "type": "bibr", "rid": "R2", "data": [ { "name": "text", "data": "2" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。由于阿尔茨海默症的发病机制尚不清楚，因此至今没有有效的药物能够彻底地根除这一病症，只能对该疾病的前期和中期的症状实现一定的抑制和缓解治疗效果。β‑淀粉样蛋白（Aβ）作为阿尔茨海默症病理性标志物之一，与AD的发病之间有着密切的关系。因此，对于Aβ蛋白的检测对AD的诊疗具有重要的意义。目前用于Aβ检测的手段主要有：正电子发射断层扫描成像（PET）、磁共振成像（MRI）、光声成像（PA）以及荧光成像（FI）等。相对于其他成像方法来说，荧光成像方法具有设备低廉、检测方便、响应快速、灵敏度高等特点" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "blockXref", "data": { "data": [ { "name": "xref", "data": { "text": "3", "type": "bibr", "rid": "R3", "data": [ { "name": "text", "data": "3" } ] } }, { "name": "text", "data": "-" }, { "name": "xref", "data": { "text": "6", "type": "bibr", "rid": "R6", "data": [ { "name": "text", "data": "6" } ] } } ], "rid": [ "R3", "R4", "R5", "R6" ], "text": "3-6", "type": "bibr" } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "，因此被广泛应用于蛋白检测。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "近几年来，关于Aβ聚集体的检测性荧光探针报道较多。最主要的分为苯并噻唑类荧光探针" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "blockXref", "data": { "data": [ { "name": "xref", "data": { "text": "7", "type": "bibr", "rid": "R7", "data": [ { "name": "text", "data": "7" } ] } }, { "name": "text", "data": "-" }, { "name": "xref", "data": { "text": "9", "type": "bibr", "rid": "R9", "data": [ { "name": "text", "data": "9" } ] } } ], "rid": [ "R7", "R8", "R9" ], "text": "7-9", "type": "bibr" } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "、姜黄素类荧光探针" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "blockXref", "data": { "data": [ { "name": "xref", "data": { "text": "10", "type": "bibr", "rid": "R10", "data": [ { "name": "text", "data": "10" } ] } }, { "name": "text", "data": "-" }, { "name": "xref", "data": { "text": "15", "type": "bibr", "rid": "R15", "data": [ { "name": "text", "data": "15" } ] } } ], "rid": [ "R10", "R11", "R12", "R13", "R14", "R15" ], "text": "10-15", "type": "bibr" } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "、BODIPY类荧光探针、花菁类荧光探针" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "blockXref", "data": { "data": [ { "name": "xref", "data": { "text": "16", "type": "bibr", "rid": "R16", "data": [ { "name": "text", "data": "16" } ] } }, { "name": "text", "data": "-" }, { "name": "xref", "data": { "text": "19", "type": "bibr", "rid": "R19", "data": [ { "name": "text", "data": "19" } ] } } ], "rid": [ "R16", "R17", "R18", "R19" ], "text": "16-19", "type": "bibr" } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "、香豆素类荧光探针" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "blockXref", "data": { "data": [ { "name": "xref", "data": { "text": "20", "type": "bibr", "rid": "R20", "data": [ { "name": "text", "data": "20" } ] } }, { "name": "text", "data": "-" }, { "name": "xref", "data": { "text": "21", "type": "bibr", "rid": "R21", "data": [ { "name": "text", "data": "21" } ] } } ], "rid": [ "R20", "R21" ], "text": "20-21", "type": "bibr" } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "以及喹啉二腈类荧光探针" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "blockXref", "data": { "data": [ { "name": "xref", "data": { "text": "22", "type": "bibr", "rid": "R22", "data": [ { "name": "text", "data": "22" } ] } }, { "name": "text", "data": "-" }, { "name": "xref", "data": { "text": "24", "type": "bibr", "rid": "R24", "data": [ { "name": "text", "data": "24" } ] } } ], "rid": [ "R22", "R23", "R24" ], "text": "22-24", "type": "bibr" } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "等。这些探针均具有良好的荧光发射波长以及较强的蛋白结合能力，但是大部分用于检测Aβ42聚集体，而用于检测Aβ40聚集体的荧光探针报道则比较少。Aβ40在AD早期患者大脑中是含量最多的一种Aβ物种，而且在AD的发病过程中，Aβ40聚集体发挥着重要作用" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "25", "type": "bibr", "rid": "R25", "data": [ { "name": "text", "data": "25" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。因此，能够实现对Aβ40聚集体的特异性检测就显得非常重要。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "本工作中基于甲氧基喹啉结构设计的荧光探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "，能够实现对Aβ40聚集体的特异性检测，该探针对Aβ40展现出良好的选择性和结合能力（" }, { "name": "italic", "data": [ { "name": "text", "data": "K" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "d" } ] }, { "name": "text", "data": "=3.072 μmol/L）。除此之外，该探针与Aβ40聚集体响应速度快，与目标蛋白结合后几秒钟就可发出强烈的荧光，并且能够保持长时间的荧光稳定性，具有在生物组织成像上的良好应用前景。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2　实　　验" } ], "level": "1", "id": "s2" } }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.1　试剂和仪器" } ], "level": "2", "id": "s2a" } }, { "name": "p", "data": [ { "name": "text", "data": "实验过程中用到的各种原料药品均为分析纯，购买自上海百灵威化学技术有限公司，纯化用的有机溶剂均为分析纯，购买自国药集团药业 股份有限公司。液相核磁共振波谱仪AV-600 NMR，AV-400 NMR，Bruker；高分辨率质谱仪MicroTOF10257，Bruker；荧光分光光度计Hitachi F-4600，日本日立高新技术公司；紫外-可见吸收光谱仪UH-5300，上海森信实验仪器有限公司；冷冻干燥机SCIENTZ-10N，宁波新芝生物科技有限公司。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.2　探针C‑1的合成及表征" } ], "level": "2", "id": "s2b" } }, { "name": "p", "data": [ { "name": "xref", "data": { "text": "图1", "type": "fig", "rid": "F1", "data": [ { "name": "text", "data": "图1" } ] } }, { "name": "text", "data": "为探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C‑1" } ] }, { "name": "text", "data": "的合成路线，具体合成方法及表征数据如下。" } ] }, { "name": "fig", "data": { "id": "F1", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图1" } ], "title": [ { "name": "text", "data": "探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C‑1" } ] }, { "name": "text", "data": "的合成路线" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig.1" } ], "title": [ { "name": "text", "data": "Synthetic route for the probe " }, { "name": "bold", "data": [ { "name": "text", "data": "C‑1" } ] } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471880&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471894&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471883&type=", "width": "139.14471436", "height": "49.43902588", "fontsize": "" } ] } }, { "name": "p", "data": [ { "name": "text", "data": "化合物" }, { "name": "bold", "data": [ { "name": "text", "data": "KL‑1" } ] }, { "name": "text", "data": "的合成：称取6-甲氧基-2-甲基喹啉（1.73 g, 10 mmol）溶解到10 mL乙腈溶液中，再加入碘甲烷（2.13 g，15 mmol），加热回流8 h，待冷却到室温，观察到有白色固体析出，抽滤得到产物" }, { "name": "bold", "data": [ { "name": "text", "data": "KL‑1" } ] }, { "name": "text", "data": "（白色固体，94.6%）。" }, { "name": "sup", "data": [ { "name": "text", "data": "1" } ] }, { "name": "text", "data": "H NMR（400 MHz，DMSO） " }, { "name": "italic", "data": [ { "name": "text", "data": "δ " } ] }, { "name": "text", "data": "= 8.94（d," }, { "name": "italic", "data": [ { "name": "text", "data": "J " } ] }, { "name": "text", "data": "= 8.6 Hz,1H）,8.51（d," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 9.4 Hz,1H）,8.05（d," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 8.6 Hz,1H）,7.84（dt," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 5.1,2.8 Hz,2H）,4.42（s,3H）,4.00（s,3H）,3.02（s,3H）。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "化合物" }, { "name": "bold", "data": [ { "name": "text", "data": "NH‑1" } ] }, { "name": "text", "data": "的合成：称取6-羟基-2-萘甲醛（0.86 g，5 mmol）和焦亚硫酸钠（1.90 g，10 mmol）于100 mL的圆底烧瓶中，加入30 mL的蒸馏水作为溶剂，再加入二甲胺水溶液10 mL，140 ℃条件下加热回流反应72 h，待到反应结束后冷却至室温，饱和食盐水洗涤，用乙酸乙酯萃取粗产物，加入无水硫酸钠除水，旋干后上样进行柱纯化，用乙酸乙酯/石油醚=1∶10做洗脱剂，得到纯的产物" }, { "name": "bold", "data": [ { "name": "text", "data": "NH‑1" } ] }, { "name": "text", "data": "（黄绿色，12.5%）。" }, { "name": "sup", "data": [ { "name": "text", "data": "1" } ] }, { "name": "text", "data": "H NMR（400 MHz,CDCl" }, { "name": "sub", "data": [ { "name": "text", "data": "3" } ] }, { "name": "text", "data": "） " }, { "name": "italic", "data": [ { "name": "text", "data": "δ" } ] }, { "name": "text", "data": " = 10.03（s,1H）,8.17（s,1H）,7.88～7.85（m,1H）,7.84（d," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 2.0 Hz,1H）,7.69（d," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 8.6 Hz,1H）,7.20（dd," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 9.1,2.6 Hz,1H）,6.91（d," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 2.5 Hz,1H）,3.15（s,6H）。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "化合物" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "的合成：称取化合物" }, { "name": "bold", "data": [ { "name": "text", "data": "NH⁃1" } ] }, { "name": "text", "data": "（39.8 mg，0.2 mmol）和化合物" }, { "name": "bold", "data": [ { "name": "text", "data": "KL⁃1" } ] }, { "name": "text", "data": "（63.0 mg，0.2 mmol）加入到无水乙醇溶液中，再加入几滴哌啶，回流反应8 h，待反应冷却后用甲醇/二氯甲烷=1∶50作洗脱剂，进行柱纯化，旋转蒸发后得到产物" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "（红色固体，28.3%）。" }, { "name": "sup", "data": [ { "name": "text", "data": "1" } ] }, { "name": "text", "data": "H NMR（400 MHz,DMSO） " }, { "name": "italic", "data": [ { "name": "text", "data": "δ" } ] }, { "name": "text", "data": " = 8.72（d," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 8.9 Hz,1H）,8.37（dd," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 18.6,9.2 Hz,2H）,8.17～8.04（m,2H）,7.95（d," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 8.4 Hz,1H）,7.69（dt," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 18.2,8.7 Hz,5H）,7.14（d," }, { "name": "italic", "data": [ { "name": "text", "data": "J" } ] }, { "name": "text", "data": " = 10.2 Hz,1H）,6.81（s,1H）,4.43（s,3H）,3.97（s,3H）,3.02 （s,6H）。" }, { "name": "sup", "data": [ { "name": "text", "data": "13" } ] }, { "name": "text", "data": "C NMR（150 MHz,DMSO） " }, { "name": "italic", "data": [ { "name": "text", "data": "δ" } ] }, { "name": "text", "data": "=158.91,153.96,150.20, 147.03,142.27,136.77,134.92,132.01,130.35,129.73,128.71,127.07,126.01,125.66,125.05,121.48,121.33,116.81,116.46,109.01, 105.66, 56.64, 55.38。HRMS（ESI," }, { "name": "italic", "data": [ { "name": "text", "data": "m" } ] }, { "name": "text", "data": "/" }, { "name": "italic", "data": [ { "name": "text", "data": "z" } ] }, { "name": "text", "data": "）:calcd for C" }, { "name": "sub", "data": [ { "name": "text", "data": "25" } ] }, { "name": "text", "data": "H" }, { "name": "sub", "data": [ { "name": "text", "data": "25" } ] }, { "name": "text", "data": "N" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sup", "data": [ { "name": "text", "data": "+" } ] }, { "name": "text", "data": ", 369.196 1; f ound:369.196 7。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.3　各种蛋白的制备" } ], "level": "2", "id": "s2c" } }, { "name": "p", "data": [ { "name": "text", "data": "Aβ40单体的制备：称取Aβ40单体蛋白1 mg，溶解于2 mL pH=7.4的磷酸缓冲溶液（PBS）中，摇晃均匀，配制成为100 μmol/L的溶液，放入冰箱冷冻备用。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "Aβ42单体的制备：称取Aβ42单体蛋白1 mg，溶解于2 mL PBS溶液中，摇晃均匀，配制成为100 μmol/L的溶液，放入冰箱冷冻备用。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "VQIVYK短肽蛋白的制备：称取VQIVYK短肽蛋白1 mg，溶解于2 mL的PBS溶液中，再加入1.44 mg的肝素钠，磁力搅拌2~3 d，配制成为600 μmol/L的溶液，放入冰箱冷冻备用。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "Aβ40聚集体的制备：称取Aβ40单体蛋白1 mg，溶解于2 mL PBS溶液中，磁力搅拌2~3 d，配制成为100 μmol/L的溶液，放入冰箱冷冻备用。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "Aβ42聚集体的制备：称取Aβ42单体蛋白1 mg，溶解于2 mL PBS溶液中，磁力搅拌2~3 d，配制成为100 μmol/L的溶液，放入冰箱冷冻备用。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "Amylin蛋白的制备：类似于Aβ蛋白的制备，取0.78 mg溶解于2 mL PBS溶液中，磁力搅拌2~3 d，配制成为100 μmol/L的溶液，放入冰箱备用。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.4　活性物质的制备" } ], "level": "2", "id": "s2d" } }, { "name": "p", "data": [ { "name": "text", "data": "ROS/RNS溶液的配制：取20.24 μL的30% H" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "溶液定容至10 mL去离子水中，配制成为20 mmol/L的H" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "溶液。取有效氯含量为5%的NaClO溶液10 μL定容于2 mL去离子水中，配制成为5 mmol/L的HOCl溶液。取27.7 μL的叔丁基过氧化氢定容于10 mL的去离子水中，配制成为20 mmol/L的TBHP溶液。称取59.59 mg的亚硝基铁氰化钠定容于10 mL去离子水中，配制成为20 mmol/L的NO溶液。取54.24 mg的偶氮二异丁脒盐酸盐定容于10 mL去离子水中，配制成为20 mmol/L的ROO" }, { "name": "bold", "data": [ { "name": "text", "data": "·" } ] }, { "name": "text", "data": "溶液。用1 mmol/L的Fe" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "溶液和200 μmol/L的H" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "溶液配制成为200 μmol/L的" }, { "name": "bold", "data": [ { "name": "text", "data": "·" } ] }, { "name": "text", "data": "OH溶液。用1 mmol/L的Fe" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "溶液和200 μmol/L的TBHP溶液配制成200 μmol/L的TBO" }, { "name": "bold", "data": [ { "name": "text", "data": "·" } ] }, { "name": "text", "data": "溶液。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "阴阳离子溶液的配制：称取4 mg的无水乙酸钠定容于5 mL去离子水中，配制成为10 mmol/L的CO" }, { "name": "sub", "data": [ { "name": "text", "data": "3" } ] }, { "name": "text", "data": "COO" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "溶液。同理，称取5.3 mg的无水碳酸钠配制成CO" }, { "name": "sub", "data": [ { "name": "text", "data": "3" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "溶液。称取12.3 mg的硫酸镁配制成SO" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "溶液。称取2.9 mg的氯化钠配成Cl" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "溶液。称取2.1 mg的氟化钠配制成F" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "溶液。称取8.3 mg的碘化钾配制成I" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "溶液。称取3.4 mg的亚硝酸钠配制成NO" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "溶液。称取8.7 mg的低亚硫酸钠配制成S" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "溶液。称取8.8 mg的乙酸镍配制成Ni" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "溶液。称取10.2 mg的氯化镁配制成Mg" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "溶液。称取12.1 mg的六水氯化铝配制成Al" }, { "name": "sup", "data": [ { "name": "text", "data": "3+" } ] }, { "name": "text", "data": "溶液。称取8.8 mg的乙酸钙配制成Ca" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "溶液。称取3.7 mg的氯化钾配制成K" }, { "name": "sup", "data": [ { "name": "text", "data": "+" } ] }, { "name": "text", "data": "溶液。称取13.9 mg的七水合硫酸亚铁配制成Fe" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "溶液。称取8.6 mg的氯化铜配制成Cu" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "溶液。称取3.9 mg乙酸铵配制成NH" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "+" } ] }, { "name": "text", "data": "溶液。称取10.97 mg的乙酸锌配制成Zn" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "溶液。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "氨基酸溶液的配制：分别称取5.8 mg的L-脯氨酸、5.3 mg的L-丝氨酸、4.5 mg的丙氨酸、12.3 mg的谷氨酰胺、5.9 mg的缬氨酸、6.0 mg的苏氨酸、6.6 mg的异亮氨酸、9.1 mg的酪氨酸、7.5 mg的甲硫氨酸、10.2 mg的色氨酸、9.0 mg的苯丙氨酸、6.1 mg的半胱氨酸、7.4 mg的谷氨酸和7.3 mg的赖氨酸定容于5 mL的PBS溶液中，配制成对应的10 mmol/L氨基酸溶液。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "还原性物质溶液的配制：分别取N-乙酰半胱氨酸8.2 mg、谷胱甘肽15.4 mg、葡萄糖9.9 mg以及37%~40%的甲醛水溶液3.7 μL定容至5 mL的PBS溶液中，配制成10 mmol/L的相应溶液。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.5　结合常数（" }, { "name": "italic", "data": [ { "name": "text", "data": "K" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "d" } ] }, { "name": "text", "data": "）的测定" } ], "level": "2", "id": "s2e" } }, { "name": "p", "data": [ { "name": "text", "data": "采用固定的蛋白浓度，用不同浓度的探针对其进行荧光滴定的方法，得出相应的荧光值，再通过非线性拟合计算得出探针对目标蛋白的结合常数" }, { "name": "italic", "data": [ { "name": "text", "data": "K" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "d" } ] }, { "name": "text", "data": "值，以此来进行评估探针与蛋白的结合能力。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "2.6　检出限（LOD）的测定" } ], "level": "2", "id": "s2f" } }, { "name": "p", "data": [ { "name": "text", "data": "首先，对探针的PBS溶液进行空白测定，用荧光分光光度计连续测11次，得出方差" }, { "name": "italic", "data": [ { "name": "text", "data": "σ" } ] }, { "name": "text", "data": "，作为仪器的误差校正，再根据蛋白对探针浓度滴定的线性方程斜率" }, { "name": "italic", "data": [ { "name": "text", "data": "k" } ] }, { "name": "text", "data": "，代入检出限（LOD）计算公式" }, { "name": "italic", "data": [ { "name": "text", "data": "D" } ] }, { "name": "text", "data": "=3" }, { "name": "italic", "data": [ { "name": "text", "data": "σ/k" } ] }, { "name": "text", "data": "，计算得出检出限。" } ] } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "3　结果与讨论" } ], "level": "1", "id": "s3" } }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "3.1　选择性和结合力" } ], "level": "2", "id": "s3a" } }, { "name": "p", "data": [ { "name": "text", "data": "首先，测定了探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "的吸收、荧光激发和发射光谱（图S1），得出其光谱属性，" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "的发射波长在640 nm处。之后对该探针的性能进行测定，首先测定了" }, { "name": "bold", "data": [ { "name": "text", "data": "C‑1" } ] }, { "name": "text", "data": "对各种蛋白的选择性。选用VQIVYK短肽（Val" }, { "name": "bold", "data": [ { "name": "text", "data": "‑" } ] }, { "name": "text", "data": "Gln" }, { "name": "bold", "data": [ { "name": "text", "data": "‑" } ] }, { "name": "text", "data": "Ile" }, { "name": "bold", "data": [ { "name": "text", "data": "‑" } ] }, { "name": "text", "data": "Val" }, { "name": "bold", "data": [ { "name": "text", "data": "‑" } ] }, { "name": "text", "data": "Tyr" }, { "name": "bold", "data": [ { "name": "text", "data": "‑" } ] }, { "name": "text", "data": "Lys，又称PHF6，Tau蛋白R3序列中一段关键蛋白）、Aβ40单体、Aβ40聚集体、Aβ42单体以及Aβ42聚集体进行测试。探针的浓度为2 μmol/L（PBS溶液，pH=7.4），VQIVYK短肽浓度为50 μmol/L，其余各种蛋白的浓度均为5 μmol/L（PBS溶液，pH=7.4）。由" }, { "name": "xref", "data": { "text": "图2", "type": "fig", "rid": "F2", "data": [ { "name": "text", "data": "图2" } ] } }, { "name": "text", "data": "（a）可以明显地看出，探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C‑1" } ] }, { "name": "text", "data": "与Aβ40聚集体结合后表现出的荧光强度最强，对其他各种蛋白的响应很弱。我们测试了探针结合Aβ40前后的绝对荧光量子产率变化。未与Aβ40结合时，探针的量子产率为" }, { "name": "italic", "data": [ { "name": "text", "data": "Φ" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "C-1" } ] }, { "name": "text", "data": "=0.6%；与Aβ40结合后，其量子效率得到了较大的提升，" }, { "name": "italic", "data": [ { "name": "text", "data": "Φ" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "C-1+Aβ" } ] }, { "name": "text", "data": "=2.4%。以上结果说明探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C‑1" } ] }, { "name": "text", "data": "可以与Aβ40聚集体有良好的荧光响应和选择性。除此之外，我们还测试了探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C‑1" } ] }, { "name": "text", "data": "对人血清白蛋白以及Amylin蛋白的选择性，由图S2可以看出，人血清蛋白和Amylin蛋白同样不会对探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C‑1" } ] }, { "name": "text", "data": "的选择性造成影响。研究了探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C‑1" } ] }, { "name": "text", "data": "与Aβ40聚集体的结合能力。使用固定的蛋白浓度，采用探针对其进行荧光滴定的方法，得出相应的荧光值，再通过非线性拟合得出探针对Aβ40聚集体的结合数值，以此来评估探针与蛋白的结合能力。由" }, { "name": "xref", "data": { "text": "图2", "type": "fig", "rid": "F2", "data": [ { "name": "text", "data": "图2" } ] } }, { "name": "text", "data": "（b）可见，结合常数" }, { "name": "italic", "data": [ { "name": "text", "data": "K" } ] }, { "name": "sub", "data": [ { "name": "text", "data": "d" } ] }, { "name": "text", "data": "=3.072 μmol/L，" }, { "name": "italic", "data": [ { "name": "text", "data": "R" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "=0.973 2，探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "与Aβ40聚集体有着较强的结合能力。进一步探究探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "与Aβ40聚集体浓度的关系，由" }, { "name": "xref", "data": { "text": "图2", "type": "fig", "rid": "F2", "data": [ { "name": "text", "data": "图2" } ] } }, { "name": "text", "data": "（c）可见，固定探针浓度为2 μmol/L，用Aβ40聚集体对探针进行浓度滴定。随着Aβ40聚集体浓度由0～7 μmol/L逐渐增加，探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "的荧光强度也随之逐渐增强，且强度增幅较大，使探针能够对目标蛋白的定性检测更加明显。通过对荧光数值进行定量的数据处理，能够得出探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "与Aβ40聚集体结合的线性关系， " }, { "name": "italic", "data": [ { "name": "text", "data": "R" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "=0.96（" }, { "name": "xref", "data": { "text": "图2", "type": "fig", "rid": "F2", "data": [ { "name": "text", "data": "图2" } ] } }, { "name": "text", "data": "（d））。除此之外，探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "表现出较高的灵敏度，检出限" }, { "name": "italic", "data": [ { "name": "text", "data": "D" } ] }, { "name": "text", "data": "=2.77 μmol/L。" } ] }, { "name": "fig", "data": { "id": "F2", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图2" } ], "title": [ { "name": "text", "data": "探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "对Aβ40的响应特性。 （a）" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "对各种蛋白的选择性；（b）" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "的结合常数；（c）Aβ40对" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "的浓度滴定；（d）" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "与Aβ40浓度的线性关系以及检出限。" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig. 2" } ], "title": [ { "name": "text", "data": "Response characteristics of probe " }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": " to Aβ40. （a）Selectivity of " }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1 " } ] }, { "name": "text", "data": "for proteins. （b）Binding constant of " }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": ". （c）Concentration titration of Aβ40 to " }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": ". （d）Linearity of " }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": " and Aβ40 concentrations and detection limits." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471898&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471909&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471902&type=", "width": "121.41754150", "height": "86.29809570", "fontsize": "" } ] } } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "3.2　抗干扰性" } ], "level": "2", "id": "s3b" } }, { "name": "p", "data": [ { "name": "text", "data": "荧光探针的亮度和稳定性是决定荧光成像技术的灵敏度和检测限的重要因素" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "26", "type": "bibr", "rid": "R26", "data": [ { "name": "text", "data": "26" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "。因此，我们研究了探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "的抗干扰稳定性测试，分别测试了探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "在多种氨基酸、RNS/ROS、各种阴阳离子以及还原性物质存在下的抗干扰能力。首先测定探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "与Aβ40聚集体结合后在多种氨基酸存在下的抗干扰性，在2 μmol/L（PBS溶液，pH=7.4）的探针和5 μmol/L（PBS溶液，pH=7.4）的Aβ40聚集体存在的条件下，分别加入5 μmol/L（PBS溶液，pH=7.4）的L-脯氨酸（Pro）、L-丝氨酸（Ser）、丙氨酸（Ala）、谷氨酰胺（Gln）、缬氨酸（Val）、苏氨酸（Thr）、异亮氨酸（Ile）、酪氨酸（Tyr）、甲硫氨酸（Met）、色氨酸（Trp）、苯丙氨酸（Phe）、半胱氨酸（Cys）、谷氨酸（Glu）和赖氨酸（Lys）。通过荧光分光光度计测定荧光强度变化，在多种氨基酸存在下，Aβ40聚集体与探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "结合后的荧光强度无明显变化，各种氨基酸的存在不会对探针的检测作用造成干扰（" }, { "name": "xref", "data": { "text": "图3", "type": "fig", "rid": "F3", "data": [ { "name": "text", "data": "图3" } ] } }, { "name": "text", "data": "（a））。" } ] }, { "name": "fig", "data": { "id": "F3", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图3" } ], "title": [ { "name": "text", "data": "探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "的抗干扰性测试。 （a）A～P：探针、Aβ40、Ala、Cys、Gln、Glu、Ile、Lys、Met、Phe、Pro、Ser、Thr、Trp、Tyr、Val；（b）A～I：探针、Aβ40、ROO" }, { "name": "bold", "data": [ { "name": "text", "data": "·" } ] }, { "name": "text", "data": "、HOCl、H" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "、TBO·、TBHP、NO、" }, { "name": "bold", "data": [ { "name": "text", "data": "·" } ] }, { "name": "text", "data": "OH；（c）A～I：探针、Aβ40、Al" }, { "name": "sup", "data": [ { "name": "text", "data": "3+" } ] }, { "name": "text", "data": "、Ca" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "、Fe" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "、Zn" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "、K" }, { "name": "sup", "data": [ { "name": "text", "data": "+" } ] }, { "name": "text", "data": "、Mg" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "、NH" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "+" } ] }, { "name": "text", "data": "、Ni" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "；（d）A～J：探针、Aβ40、Cl" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、CO" }, { "name": "sub", "data": [ { "name": "text", "data": "3" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "、CO" }, { "name": "sub", "data": [ { "name": "text", "data": "3" } ] }, { "name": "text", "data": "COO" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、F" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、I" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、NO" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、S" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "、SO" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "。" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig. 3" } ], "title": [ { "name": "text", "data": "Interference resistance of the probe " }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": ". （a）A-P： probe，Aβ40，Ala，Cys，Gln，Glu，Ile，Lys，Met，Phe，Pro，Ser，Thr，Trp，Tyr，Val. （b）A-I： probe，Aβ40，ROO" }, { "name": "bold", "data": [ { "name": "text", "data": "·，" } ] }, { "name": "text", "data": "HOCl，H" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "，TBO·，TBHP，NO，" }, { "name": "bold", "data": [ { "name": "text", "data": "·" } ] }, { "name": "text", "data": "OH. （c）A-I： probe，Aβ40， Al" }, { "name": "sup", "data": [ { "name": "text", "data": "3+" } ] }, { "name": "text", "data": "，Ca" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "，Fe" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "，Zn" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "， K" }, { "name": "sup", "data": [ { "name": "text", "data": "+" } ] }, { "name": "text", "data": "， Mg" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "， NH" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "+" } ] }, { "name": "text", "data": "， Ni" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": ". （d）A-J： probe， Aβ40， Cl" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "， CO" }, { "name": "sub", "data": [ { "name": "text", "data": "3" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "， CO" }, { "name": "sub", "data": [ { "name": "text", "data": "3" } ] }, { "name": "text", "data": "COO" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "， F" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "， I" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "， NO" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "， S" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "， SO" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471914&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471922&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471919&type=", "width": "119.92669678", "height": "86.41464233", "fontsize": "" } ] } }, { "name": "p", "data": [ { "name": "text", "data": "选用ROO" }, { "name": "bold", "data": [ { "name": "text", "data": "·" } ] }, { "name": "text", "data": "、HOCl、H" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "、TBO" }, { "name": "bold", "data": [ { "name": "text", "data": "·" } ] }, { "name": "text", "data": "、TBHP、NO以及" }, { "name": "bold", "data": [ { "name": "text", "data": "·" } ] }, { "name": "text", "data": "OH等多种ROS/RNS来测定荧光探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "在该类活性物质存在条件下的抗干扰性。探针浓度2 μmol/L（PBS溶液，pH=7.4），Aβ40聚集体和各种活性物质的浓度均为5 μmol/L（PBS溶液，pH=7.4）,进行相关的实验测试。由" }, { "name": "xref", "data": { "text": "图3", "type": "fig", "rid": "F3", "data": [ { "name": "text", "data": "图3" } ] } }, { "name": "text", "data": "（b）可见，探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "在多种活性氧和活性氮物质存在下，其荧光强度仍然能够维持一个稳定的状态，说明探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "的荧光成像不受该类物质的干扰。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "然后，我们测定了探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "在不同阴阳离子存在条件下的抗干扰能力。分别选用Al" }, { "name": "sup", "data": [ { "name": "text", "data": "3+" } ] }, { "name": "text", "data": "、Ca" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "、Fe" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "、Zn" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "、K" }, { "name": "sup", "data": [ { "name": "text", "data": "+" } ] }, { "name": "text", "data": "、Mg" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "、NH" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "+" } ] }, { "name": "text", "data": "和Ni" }, { "name": "sup", "data": [ { "name": "text", "data": "2+" } ] }, { "name": "text", "data": "等阳离子（" }, { "name": "xref", "data": { "text": "图3", "type": "fig", "rid": "F3", "data": [ { "name": "text", "data": "图3" } ] } }, { "name": "text", "data": "（c））,以及Cl" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、CO" }, { "name": "sub", "data": [ { "name": "text", "data": "3" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "、CO" }, { "name": "sub", "data": [ { "name": "text", "data": "3" } ] }, { "name": "text", "data": "COO" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、F" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、I" }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、NO" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "-" } ] }, { "name": "text", "data": "、S" }, { "name": "sub", "data": [ { "name": "text", "data": "2" } ] }, { "name": "text", "data": "O" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "和SO" }, { "name": "sub", "data": [ { "name": "text", "data": "4" } ] }, { "name": "sup", "data": [ { "name": "text", "data": "2-" } ] }, { "name": "text", "data": "等阴离子（" }, { "name": "xref", "data": { "text": "图3", "type": "fig", "rid": "F3", "data": [ { "name": "text", "data": "图3" } ] } }, { "name": "text", "data": "（d））来测定探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "在生物环境溶液中的抗干扰能力。探针的浓度为2 μmol/L（PBS溶液，pH=7.4），目标蛋白和各种离子的浓度均为5 μmol/L（PBS溶液，pH=7.4）。实验结果表明，探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "同样表现出了良好的抗干扰性。" } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "3.3　稳定性" } ], "level": "2", "id": "s3c" } }, { "name": "p", "data": [ { "name": "text", "data": "接下来，我们测定了探针在谷胱甘肽（GSH）、N-乙酰-L-半胱氨酸（NAC）、甲醛（HCHO）以及葡萄糖（Glucose）四种还原性物质存在下的稳定性。探针浓度为2 μmol/L（PBS溶液，pH=7.4），目标蛋白的浓度为5 μmol/L（PBS溶液，pH=7.4），还原性物质的用量分别为目标蛋白的0，1，2.5，5，10个当量。探针 " }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "在谷胱甘肽（" }, { "name": "xref", "data": { "text": "图4", "type": "fig", "rid": "F4", "data": [ { "name": "text", "data": "图4" } ] } }, { "name": "text", "data": "（a））、NAC（" }, { "name": "xref", "data": { "text": "图4", "type": "fig", "rid": "F4", "data": [ { "name": "text", "data": "图4" } ] } }, { "name": "text", "data": "（b））、甲醛（" }, { "name": "xref", "data": { "text": "图 4", "type": "fig", "rid": "F4", "data": [ { "name": "text", "data": "图 4" } ] } }, { "name": "text", "data": "（c））以及Glucose（" }, { "name": "xref", "data": { "text": "图4", "type": "fig", "rid": "F4", "data": [ { "name": "text", "data": "图4" } ] } }, { "name": "text", "data": "（d））的0~10个当量存在条件下，依旧能够与Aβ40聚集体结合且保持良好的荧光稳定。探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "在生物体系溶液环境中，完全不受多种还原性物质的干扰。" } ] }, { "name": "fig", "data": { "id": "F4", "caption": [ { "lang": "zh", "label": [ { "name": "text", "data": "图4" } ], "title": [ { "name": "text", "data": "探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "与Aβ40在GSH（a）、NAC（b）、HCHO（c）和葡萄糖（d）存在情况下的稳定性测试。" } ] }, { "lang": "en", "label": [ { "name": "text", "data": "Fig. 4" } ], "title": [ { "name": "text", "data": "Stability of the probe " }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": " with Aβ40 in the presence of GSH（a）， NAC（b）， HCHO（c） and Glucose（d）." } ] } ], "subcaption": [], "note": [], "graphics": [ { "print": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471929&type=", "small": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471940&type=", "big": "http://html.publish.founderss.cn/rc-pub/api/common/picture?pictureId=36471935&type=", "width": "124.65313721", "height": "91.79815674", "fontsize": "" } ] } }, { "name": "p", "data": [ { "name": "text", "data": "我们还测定了探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "随时间变化以及在不同pH值下的稳定性。由图S3（a）可以看出，探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "与Aβ40响应极快，加入后几秒钟就可发出强荧光，且随时间延长能够保持较强的稳定性，经过30 min后仍然能保持荧光稳定。配制pH值范围为2~11的磷酸缓冲溶液。由图S3（b）可以清晰地看出，无论是在酸性或者是碱性条件下，探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "自身以及与Aβ40结合后，均能够保持荧光稳定性。最后，我们测定了该探针的生物相容性，通过MTT法" }, { "name": "sup", "data": [ { "name": "text", "data": "[" }, { "name": "xref", "data": { "text": "27", "type": "bibr", "rid": "R27", "data": [ { "name": "text", "data": "27" } ] } }, { "name": "text", "data": "]" } ] }, { "name": "text", "data": "进行评估。选用Hela细胞在37 ℃条件下与探针共同孵育24 h，由图S4可以看出，即使探针的浓度达到了40 μmol/L，细胞的存活率仍然能够达到80%以上，证明了该探针的低毒性。" } ] } ] } ] }, { "name": "sec", "data": [ { "name": "sectitle", "data": { "title": [ { "name": "text", "data": "4　结　　论" } ], "level": "1", "id": "s4" } }, { "name": "p", "data": [ { "name": "text", "data": "在本工作中，我们设计合成了近红外荧光探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "，实现了对Aβ40聚集体良好的特异性检测。探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "具有640 nm的荧光发射波长，且与Aβ40聚集体的结合能力强，响应速度快，抗干扰能力强，稳定性好，并且背景干扰小，在溶液测试中表现出较好的性能。除此之外，该探针的生物相容性良好。综上所述，探针" }, { "name": "bold", "data": [ { "name": "text", "data": "C⁃1" } ] }, { "name": "text", "data": "具备的这些优点可能会使其在生物成像上具有一定的应用潜力。" } ] }, { "name": "p", "data": [ { "name": "text", "data": "本文补充文件和专家审稿意见及作者回复内容的下载地址：" }, { "name": "extlink", "data": { "text": [ { "name": "text", "data": "http://cjl.lightpublishing.cn/thesisDetails#10.37188/CJL.20220228." } ], "href": "http://cjl.lightpublishing.cn/thesisDetails#10.37188/CJL.20220228." } } ] } ] } ], "footnote": [], "reflist": { "title": [ { "name": "text", "data": "参 考 文 献" } ], "data": [ { "id": "R1", "label": "1", "citation": [ { "lang": "en", "text": [ { "name": "text", "data": "NASICA-LABOUZE J" }, { "name": "text", "data": "， " }, { "name": "text", "data": "NGUYEN P H" }, { "name": "text", "data": "， " }, { "name": "text", "data": "STERPONE F" }, { "name": "text", "data": "， " }, { "name": "text", "data": "et al" }, { "name": 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