Xing Hu, Zhang Yiting, Li Runliu, Ruzicka Hans-Maximilian, Hain Christopher, Andersson Jakob, Bozdogan Anil, Henkel Marius, Knippschild Uwe, Hasler Roger, Kleber Christoph, Knoll Wolfgang, Kissmann Ann-Kathrin, Rosenau Frank
Institute of Pharmaceutical Biotechnology, Ulm University, Albert-Einstein-Allee 11, 89081 Ulm, Germany.
AIT Austrian Institute of Technology GmbH, Giefinggasse 4, 1210 Vienna, Austria.
Nanoscale Horiz. 2024 Dec 16;10(1):124-134. doi: 10.1039/d4nh00281d.
The use of health-relevant bacteria originating from human microbiomes for the control or therapy of diseases, including neurodegenerative disorders or diabetes, is currently gaining increasing importance in medicine. Directed and successful engineering of microbiomes probiotic supplementation requires subtle, precise as well as, more importantly, easy, fast and convenient monitoring of its success, , in patients' gut. Based on a previously described polyclonal SELEX aptamer library evolved against the human gut bacterium , we finally isolated three individual aptamers that proved their performance concerning affinity, specificity and robustness in reliably labeling the target bacterium and in combination with "contaminating" control bacteria. Using biofunctionalization molecules on gFETs, we could specifically quantify 10-10 cells per mL, retrace their number in mixtures and determine aptamer -values around 2 nM. These measurements were possible even in the context of a real human stool sample. Our results qualify gFETs in combination with BL2, BL7 and BL8 aptamers as a promising foundation for the construction of respective sensing devices, which will open new avenues towards developing an intended monitoring technique for probiotic therapy and microbiome engineering approaches.
源自人类微生物群的与健康相关的细菌用于疾病(包括神经退行性疾病或糖尿病)的控制或治疗,目前在医学上正变得越来越重要。微生物群的定向且成功的工程改造——益生菌补充,需要对其在患者肠道中的成功情况进行微妙、精确且更重要的是简单、快速和方便的监测。基于先前描述的针对人类肠道细菌进化的多克隆SELEX适配体文库,我们最终分离出三种单独的适配体,它们在可靠标记目标细菌以及与“污染”对照细菌结合方面,在亲和力、特异性和稳健性方面证明了其性能。使用基于石墨烯场效应晶体管(gFET)的生物功能化分子,我们能够特异性地定量每毫升10-10个细胞,追踪它们在混合物中的数量,并确定适配体的解离常数(Kd)值约为2 nM。即使在真实的人类粪便样本的情况下,这些测量也是可行的。我们的结果使gFET与BL2、BL7和BL8适配体相结合,成为构建相应传感设备的有前途的基础,这将为开发用于益生菌治疗和微生物群工程方法的预期监测技术开辟新途径。