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Single-Molecule Identification of the Conformations of Human C-Reactive Protein and Its Aptamer Complex with Solid-State Nanopores
Wu, Ji1,2; Liang, Liyuan1,2; Zhang, Mingkun1,2; Zhu, Rui1,2; Wang, Zhong1,2; Yin, Yajie1,2; Yin, Bohua1,2; Weng, Ting1,2; Fang, Shaoxi1,2; Xie, Wanyi1,2
2022-03-16
摘要Human C-reactive protein (CRP) is an established inflammatory biomarker and was proved to be potentially relevant to disease pathology and cancer progression. A large body of methodologies have been reported for CRP analysis, including electrochemical/optical biosensors, aptamer, or antibody-based detection. Although the detection limit is rather low until pg/uL, most of which are time-consuming and relatively expensive, and few of them provided CRP single-molecule information. This work demonstrated the nanopore-based approach for the characterization of CRP conformation under versatile conditions. With an optimized pore of 14 nm in diameter, we achieved the detection limit as low as 0.3 ng/mu L, voltage polarity significantly influences the electro-osmotic force and CRP translocation behavior, and the pentameric conformation of CRP may dissociate into pro-inflammatory CRP isoforms and monomeric CRP at bias potential above 300 mV. CRP tends to translocate through nanopores faster along with the increase in pH values, due to more surface charge on both CRP and pore inner wall and stronger electro-osmotic force. The CRP could specifically bind with its aptamer of different concentrations to form complexes, and the complexes exhibited distinguishable nanopore translocation behavior compared with CRP alone. The variation of the molar ratio of aptamer significantly influences the orientation of CRP translocation. The plasma test under physiological conditions displayed the ability of the nanopore system on the CRP identification with a concentration of 3 ng/mu L.
关键词solid-state nanopore C-reactive protein aptamer single-molecule conformation
DOI10.1021/acsami.2c00453
发表期刊ACS APPLIED MATERIALS & INTERFACES
ISSN1944-8244
卷号14期号:10页码:12077-12088
通讯作者Liang, Liyuan(liangliyuan@cigit.ac.cn)
收录类别SCI
WOS记录号WOS:000787549000006
语种英语