水溶性可见光激发铕荧光生物标记物的制备与应用  

Preparation and application of a water-soluble visible-light-sensitized luminescent europium biolabel

作　　者：蒋丽娜[1] 叶志强[1] 王桂兰[1] 袁景利[1] 

机构地区：[1]大连理工大学化学系精细化工国家重点实验室,辽宁大连116012

出　　处：《中国科技论文在线》2009年第12期915-918,共4页

基　　金：国家自然科学基金(20835001);高等学校博士学科点专项科研基金(200801410003)

摘　　要：大多数稀土荧光生物标记物需使用光毒性较大的紫外光作为激发光源,对活体生物样品有一定的光损伤,而已知的几种长波长激发铕配合物又存在水溶性差、极性配位溶剂中不稳定、发光量子产率低及缺乏活性标记基团等问题而无法用于生物标记。通过将一种可见光激发铕(Ⅲ)配合物与牛血清白蛋白结合的方法,制备出了水溶性好、激发波长延长至可见光区(<460nm)并具有较高荧光量子产率(27%)和较长荧光寿命(0.46ms)的铕(Ⅲ)配合物结合蛋白。该结合蛋白标记链霉亲和素后被成功地用于水样品中病原微生物小隐孢子虫(Cryptosporidium parvum)的免疫荧光染色及时间分辨荧光成像测定。For most luminescent lanthainde biolabels,one of the major drawbacks is that the optical excitation window is limited to the UV region,which has stronger phototoxicity on the living biological samples. Although the excitation wavelengths for several Eu^3＋ complexes can be extended into the visible region,water insolubility,poor stability in polar solvents,low luminescence quantum yield,and the lack of active groups of these complexes make them cannot be used for biolabeling. In this work,by conjugating bovine serum albumin （BSA） with a visible-light-sensitized Eu^3＋ complex,a Eu^3＋ complex-BSA conjugate ,which has good water solubility,visible-light excitability （460 nm）,longer luminescence lifetime （〈0.46 ms）,and larger quantum yield （27%） was prepared. The Eu^3＋ complex-BSA conjugate-labeled streptavidin was perpared,and successfully used for immuno-staining and time-resolved luminescence imaging detection of an environmental pathogen,Cryptosporidium parvum,in the water sample.

关 键 词：铕配合物 生物标记 环境病原微生物 时间分辨荧光 生物成像 

分 类 号：Q503[生物学—生物化学]