Liu Yang, Danielsson Bengt
Pure and Applied Biochemistry, Lund University, P.O. Box 124, SE-22100 Lund, Sweden.
Anal Chim Acta. 2007 Mar 21;587(1):47-51. doi: 10.1016/j.aca.2007.01.013. Epub 2007 Jan 16.
A double stranded DNA based fluorescence bioprobe for anticancer agent (doxorubicin) detection is described. This method provides a new way for sensitive DNA/drug interaction study by a homogeneous assay. The probe employs the long-wavelength intercalating fluorophore TOTO-3 (TT3). The anticancer agent, doxorubicin, which interacts with the DNA-TT3 complex, was indirectly measured by the decrease in the fluorescence intensity. Various oligonucleotides with different sequences were examined. Doxorubicin has preference for the oligonucleotide 5'AGCACG3'. Enhanced fluorescence observed for the TT3 intercalation with this oligonucleotide makes the DNA-dye complex a suitable bioprobe for doxorubicin detection by competitive assay. A home-built CCD camera setup was applied along with 384 well plate assay format for high throughput fluorescence imaging. The detection limit can be as low as 25 ng mL(-1) with an upper limit of 100 microg mL(-1). The recovery test with spiked serum sample shows that this method can be a potential routine method for therapeutic drug monitoring (TDM).
描述了一种用于检测抗癌剂（阿霉素）的基于双链DNA的荧光生物探针。该方法通过均相分析为敏感的DNA/药物相互作用研究提供了一种新途径。该探针采用长波长嵌入荧光团TOTO-3（TT3）。与DNA-TT3复合物相互作用的抗癌剂阿霉素通过荧光强度的降低来间接测量。研究了具有不同序列的各种寡核苷酸。阿霉素对寡核苷酸5'AGCACG3'有偏好。观察到TT3与该寡核苷酸嵌入时荧光增强，使得DNA-染料复合物成为通过竞争分析检测阿霉素的合适生物探针。将自制的电荷耦合器件（CCD）相机装置与384孔板分析形式一起应用于高通量荧光成像。检测限可低至25 ng mL(-1)，上限为100 μg mL(-1)。加标血清样品的回收率测试表明，该方法可能成为治疗药物监测（TDM）的潜在常规方法。
