Li Shanghao, Mulloor Jerome J, Wang Lingyu, Ji Yiwen, Mulloor Catherine J, Micic Miodrag, Orbulescu Jhony, Leblanc Roger M
Department of Chemistry and ‡Department of Biology, University of Miami , 1301 Memorial Drive, Cox Science Center, Coral Gables, Florida 33146, United States.
ACS Appl Mater Interfaces. 2014 Apr 23;6(8):5704-12. doi: 10.1021/am500254e. Epub 2014 Apr 8.
Biosensing methods and devices using graphene oxide (GO) have recently been explored for detection and quantification of specific biomolecules from body fluid samples, such as saliva, milk, urine, and serum. For a practical diagnostics application, any sensing system must show an absence of nonselective detection of abundant proteins in the fluid matrix. Because lysozyme is an abundant protein in these body fluids (e.g., around 21.4 and 7 μg/mL of lysozyme is found in human milk and saliva from healthy individuals, and more than 15 or even 100 μg/mL in patients suffering from leukemia, renal disease, and sarcoidosis), it may interfere with detections and quantification if it has strong interaction with GO. Therefore, one fundamental question that needs to be addressed before any development of GO based diagnostics method is how GO interacts with lysozyme. In this study, GO has demonstrated a strong interaction with lysozyme. This interaction is so strong that we are able to subsequently eliminate and separate lysozyme from aqueous solution onto the surface of GO. Furthermore, the strong electrostatic interaction also renders the selective adsorption of lysozyme on GO from a mixture of binary and ternary proteins. This selectivity is confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), fluorescence spectroscopy, and UV-vis absorption spectroscopy.
最近,人们探索了使用氧化石墨烯(GO)的生物传感方法和设备,用于检测和定量来自体液样本(如唾液、牛奶、尿液和血清)中的特定生物分子。对于实际的诊断应用,任何传感系统都必须在流体基质中不出现对丰富蛋白质的非选择性检测。因为溶菌酶是这些体液中的一种丰富蛋白质(例如,健康个体的人乳和唾液中溶菌酶的含量约为21.4和7μg/mL,而白血病、肾病和结节病患者的溶菌酶含量超过15甚至100μg/mL),如果它与GO有强烈相互作用,可能会干扰检测和定量。因此,在开发任何基于GO的诊断方法之前,需要解决的一个基本问题是GO如何与溶菌酶相互作用。在本研究中,GO已证明与溶菌酶有强烈相互作用。这种相互作用非常强烈,以至于我们能够随后将溶菌酶从水溶液中消除并分离到GO表面。此外,强烈的静电相互作用还使得溶菌酶能够从二元和三元蛋白质混合物中选择性吸附在GO上。这种选择性通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、荧光光谱和紫外可见吸收光谱得到证实。