Institute of Biochemistry and Biology, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476 Potsdam, Germany.
Faculty of Health Sciences Brandenburg, Brandenburg University of Technology, Cottbus-Senftenberg, Universitätsplatz 1, 01968 Senftenberg, Germany.
Int J Mol Sci. 2022 Jun 3;23(11):6275. doi: 10.3390/ijms23116275.
The discovery that certain diseases have specific miRNA signatures which correspond to disease progression opens a new biomarker category. The detection of these small non-coding RNAs is performed routinely using body fluids or tissues with real-time PCR, next-generation sequencing, or amplification-based miRNA assays. Antibody-based detection systems allow an easy onset handling compared to PCR or sequencing and can be considered as alternative methods to support miRNA diagnostic in the future. In this study, we describe the generation of a camelid heavy-chain-only antibody specifically recognizing miRNAs to establish an antibody-based detection method. The generation of nucleic acid-specific binders is a challenge. We selected camelid binders via phage display, expressed them as VHH as well as full-length antibodies, and characterized the binding to several miRNAs from a signature specific for dilated cardiomyopathy. The described workflow can be used to create miRNA-specific binders and establish antibody-based detection methods to provide an additional way to analyze disease-specific miRNA signatures.
某些疾病具有特定的 miRNA 特征,这些特征与疾病的进展相对应,这一发现开辟了一个新的生物标志物类别。这些小的非编码 RNA 的检测通常使用实时 PCR、下一代测序或基于扩增的 miRNA 检测方法进行,利用体液或组织进行。与 PCR 或测序相比,基于抗体的检测系统具有简单的起始处理,并且可以被认为是未来支持 miRNA 诊断的替代方法。在这项研究中,我们描述了生成专门识别 miRNA 的重链单域抗体(nanobody)以建立基于抗体的检测方法。生成核酸特异性结合物是一个挑战。我们通过噬菌体展示选择了骆驼科结合物,将其表达为 VHH 以及全长抗体,并对来自扩张型心肌病特征性 miRNA 签名的几种 miRNA 的结合进行了表征。所描述的工作流程可用于创建 miRNA 特异性结合物并建立基于抗体的检测方法,为分析疾病特异性 miRNA 特征提供了另一种方法。