Highly bright avidin-based affinity probes carrying multiple lanthanide chelates.
Highly bright avidin-based affinity probes carrying multiple lanthanide chelates.
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携带多种镧系元素螯合物的高亮度亲和素亲和探针。
DOI：
10.1016/j.jphotobiol.2012.07.001
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Mustaev,Arkady
中科院分区：
文献类型：
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作者：
Wirpsza,Laura;Pillai,Shyamala;Batish,Mona;Marras,Salvatore;Krasnoperov,Lev;Mustaev,Arkady
镧系离子发光具有长寿命，使得能够在时间选通模式下进行高灵敏度检测。可以通过使用附着到载体分子的多个发光标记来进一步增加灵敏度，所述载体分子可以与感兴趣的对象缀合。我们发现，多达30个镧系元素螯合物可以连接到抗生物素蛋白，创造高度明亮的结构。这些具有Eu 3+螯合物的构建体显示出增强重度改性样品的亮度的协同效应，而对于Tb 3+螯合物观察到相反的效应，从而显著降低它们的发光。通过在螯合物和蛋白质附着位点之间引入芳香族间隔物，这种不期望的Tb 3+发光体的猝灭被完全抑制。结合物的估计检测限在10−14-10− 15 M范围内。我们通过使用它们标记细菌和哺乳动物来源的活细胞，证明了新探针的高灵敏度。
Lanthanide ion luminescence has a long lifetime enabling highly sensitive detection in time-gated mode. The sensitivity can be further increased by using multiple luminescent labels attached to a carrier molecule, which can be conjugated to an object of interest. We found that up to 30 lanthanide chelates can be attached to avidin creating highly bright constructs. These constructs with Eu3+chelates display synergistic effect that enhance the brightness of heavily modified samples, while the opposite effect was observed for Tb3+chelates thereby significantly reducing their light emission. This undesirable quenching of Tb3+luminophores was completely suppressed by the introduction of an aromatic spacer between the chelate and the protein attachment site. The estimated detection limit for the conjugates is in the 10−14–10−15M range. We demonstrated a high sensitivity for the new probes by using them to label living cells of bacterial and mammalian origin.