Ersöz Arzu, Diltemiz S Emir, Ozcan A Atilir, Denizli Adil, Say Ridvan
Department of Chemistry, Anadolu University, Eskişehir, Turkey.
Biosens Bioelectron. 2008 Dec 1;24(4):742-7. doi: 10.1016/j.bios.2008.06.058. Epub 2008 Jul 12.
Recently, the 8-hydroxy-2'-deoxyguanosine (8-OHdG) has been used as a marker to determine the oxidative stress. There is no any cheap and easy determination method based on chips and sensor systems for the determination of 8-OHdG. In this study, we have proposed imprinting methods for 8-OHdG recognition and determination using methacryloylamidohistidine-platinum(II) [MAH-Pt(II)] as a new metal-chelating monomer. The study includes the solid-phase extraction (SPE) of blood sample by a new 8-OHdG imprinted sorbent and the measurement of binding interaction of 8-OHdG imprinted quartz crystal microbalance (QCM) sensor via ligand interaction. 8-OHdG imprinted sorbent has prepared by bulk polymerization of MAH-Pt(II) and N-N'-methylenebisacrylamide. 8-OHdG imprinted sensor has prepared on a QCM chip coating the thiol pretreated Au electrode. At the end of these steps, a thin molecular imprinted polymer (MIP) film for the detection of 8-OHdG has developed and analytical performance of QCM sensor which has prepared using MIP was investigated. The affinity constant (K(a)) for 8-OHdG using MAH-Pt-based thin film has determined by using the Scatchard method. The average percentage recovery of 8-OHdG from plasma samples was found as 80% by using of 8-OHdG imprinted SPE material. At the last step, 8-OHdG level in several blood plasma has been determined by this improved QCM sensor. The obtained results confirmed that the 8-OHdG level in cancer patient's blood was significantly higher than in general subjects.
最近,8-羟基-2'-脱氧鸟苷(8-OHdG)已被用作确定氧化应激的标志物。目前还没有基于芯片和传感器系统的廉价且简便的8-OHdG测定方法。在本研究中,我们提出了使用甲基丙烯酰氨基组氨酸-铂(II)[MAH-Pt(II)]作为新型金属螯合单体的8-OHdG识别和测定的印迹方法。该研究包括用新型8-OHdG印迹吸附剂对血样进行固相萃取(SPE),以及通过配体相互作用测量8-OHdG印迹石英晶体微天平(QCM)传感器的结合相互作用。8-OHdG印迹吸附剂通过MAH-Pt(II)和N-N'-亚甲基双丙烯酰胺的本体聚合制备。8-OHdG印迹传感器在涂有硫醇预处理金电极的QCM芯片上制备。在这些步骤结束时,开发了用于检测8-OHdG的薄分子印迹聚合物(MIP)膜,并研究了使用MIP制备的QCM传感器的分析性能。使用基于MAH-Pt的薄膜对8-OHdG的亲和常数(K(a))通过Scatchard方法测定。使用印迹了8-OHdG的SPE材料,发现血浆样品中8-OHdG的平均回收率为80%。在最后一步,通过这种改进的QCM传感器测定了几份血浆中的8-OHdG水平。所得结果证实,癌症患者血液中的8-OHdG水平明显高于普通受试者。