Total protein concentration quantification using nanobeads with a new highly luminescent terbium(III) complex

Cywinski Piotr J.; Pietraszkiewicz Marek; Maciejczyk Michal; Gorski Krzysztof; Hammann Tommy; Liermann Konstanze; Paulke Bernd-Reiner; Loehmannsroeben Hans-Gerd

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Total protein concentration quantification using nanobeads with a new highly luminescent terbium(III) complex

机译：使用纳米孔的总蛋白质浓度定量，具有新的高亮铽（III）复合物

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Total protein concentration (TPC) is a key parameter in many biochemical experiments and its quantification is often necessary for isolation, separation, and analysis of proteins. A sensitive and rapid nanobead-based TPC quantification assay based on Forster Resonance Energy Transfer (FRET) has been developed. A new, highly luminescent Tb(III) complex has been synthesised and applied as donor in this FRET assay with an organic dye (Cy5) as acceptor. FRET-induced changes in luminescence have been investigated both at donor and acceptor emission wavelength using time-resolved luminescence spectroscopy with time-gated detection. In the assay, the Tb(III) complex and fine-tuned polyglycidyl methacrylate (PGMA) nanobeads ensure that an improvement in sensitivity and background reduction is achieved. Using 40 nm large PGMA nanobeads loaded with the Tb(III) complex, it is possible to determine TPC down to 50 ng mL(-1) in just 10 minutes. Through specific assay components the sensitivity has been improved when compared to existing nanobead-based assays and to currently known commercial methods. Additionally, the assay is relatively insensitive to the presence of contaminants, such as non-ionic detergents commonly found in biological samples. Due to no need for any centrifugal steps, this mix-and-measure bioassay can easily be implemented into routine TPC quantification protocols in biochemical laboratories.

机译：总蛋白质浓度（TPC）是许多生物化学实验中的关键参数，其定量通常是分离，分离和分析蛋白质的必要条件。已经开发了一种基于孔谐振能量转移（FRET）的基于敏感和快速的基于纳米的TPC定量测定。已经合成了一种新的高发致发光的TB（III）络合物，并用作有机染料（CY5）作为受体作为受体的供体。使用时间分辨的发光光谱，在施主和受体发射波长中研究了发光的诱导变化，使用时间升定的发光光谱，具有时间门控检测。在测定中，Tb（III）复合物和微调的聚缩氟基甲基丙烯酸酯（PGMA）纳米甲基甲酸酯确保实现了敏感性和背景降低的改善。使用加载Tb（III）复合物的40nm大的PGMA纳米孔，只需10分钟就可以确定TPC至50ng（-1）。通过特异性测定组分，与现有的基于纳米孔的测定和当前已知的商业方法相比，敏感性已经提高。另外，测定对污染物的存在相对不敏感，例如在生物样品中常见的非离子洗涤剂。由于不需要任何离心步骤，这种混合和测量的生物测定可以容易地在生化实验室中的常规TPC定量方案中实施。

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《RSC Advances》 |2016年第116期|共6页
作者
Cywinski Piotr J.; Pietraszkiewicz Marek; Maciejczyk Michal; Gorski Krzysztof; Hammann Tommy; Liermann Konstanze; Paulke Bernd-Reiner; Loehmannsroeben Hans-Gerd;
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Fraunhofer Inst Appl Polymer Res Funct Mat &

Devices Geiselbergstr 69 D-14476 Potsdam Germany;

Polish Acad Sci Inst Phys Chem Kasprzaka 44-52 PL-01224 Warsaw Poland;

Polish Acad Sci Inst Phys Chem Kasprzaka 44-52 PL-01224 Warsaw Poland;

Polish Acad Sci Inst Phys Chem Kasprzaka 44-52 PL-01224 Warsaw Poland;

Fraunhofer Inst Appl Polymer Res Funct Mat &

Devices Geiselbergstr 69 D-14476 Potsdam Germany;

Fraunhofer Inst Appl Polymer Res Funct Mat &

Devices Geiselbergstr 69 D-14476 Potsdam Germany;

Fraunhofer Inst Appl Polymer Res Microencapsulat &

Particle Applicat Geiselbergstr 69 D-14476 Potsdam Germany;

Univ Potsdam Inst Chem Phys Chem Karl Liebknecht Str 24-25 D-14476 Potsdam Germany;

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1. Total protein concentration quantification using nanobeads with a new highly luminescent terbium(III) complex [J] . Cywinski Piotr J., Pietraszkiewicz Marek, Maciejczyk Michal, RSC Advances . 2016,第116期

机译：使用纳米孔的总蛋白质浓度定量，具有新的高亮铽（III）复合物
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Total protein concentration quantification using nanobeads with a new highly luminescent terbium(III) complex

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