首页 | 本学科首页   官方微博 | 高级检索  
     


Atomic force microscope-based single-molecule force spectroscopy of RNA unfolding
Authors:Heus Hans A  Puchner Elias M  van Vugt-Jonker Aafke J  Zimmermann Julia L  Gaub Hermann E
Affiliation:aBiomedical Diagnostics Institute (BDI), Dublin City University, Dublin 9, Ireland;bSchool of Biotechnology, Dublin City University, Dublin 9, Ireland;cNational Centre for Sensor Research (NCSR), Dublin City University, Dublin 9, Ireland;dPfizer Discovery Research, Global Biotherapeutics Technologies, Development Lab, Grangecastle Business Park, Dublin 22, Ireland
Abstract:
Over the past 10 years, a growing field of research supporting the value of myeloperoxidase (MPO) as a prognostic indicator in acute cardiac pathophysiologies has emerged. The availability of a rapid and disposable MPO detection platform would enable research clinicians to more readily assess MPO indications for guiding therapy and also facilitate clinicians at the patient interface to readily adopt MPO testing and potentially drive more informed prognoses. Here we describe the isolation of a high-affinity avian MPO-specific recombinant antibody panel using phage display. Rapid isolation of a suitable single-chain variable fragment (scFv) antibody was facilitated using a surface plasmon resonance (SPR)-based “off-rate ranking” screening process. The selected scFv was then successfully incorporated into a rapid, simple, and sensitive one-step lateral flow immunoassay (LFIA) for the detection of MPO. This “one-step” feature of the developed assay was made possible by the scFv’s strong affinity for MPO, obviating the need for sandwich signal enhancement steps. The assay’s rapid performance was also further enhanced by exploiting the intrinsic enzymatic properties of MPO in its final detection. Use of the optimized LFIA facilitated the sensitive detection of MPO in MPO-depleted serum within clinically relevant reference ranges.
Keywords:Cardiovascular disease   MPO   Phage display   Lateral flow   Point-of-care
本文献已被 ScienceDirect PubMed 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号