Journal
ANALYTICAL CHEMISTRY
Volume 91, Issue 16, Pages 10879-10886Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.9b02697
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Funding
- National Key R&D Program of China [2018YFC1602900]
- National Science Foundation of China [21735004, 21874089, 21775128, 21705024]
- Program for Changjiang Scholars and Innovative Research Team in University [IRT13036]
- Innovative research team of high-level local universities in Shanghai
- Ministry of Education [SCAI1801]
- Key Laboratory of Spectrochemical Analysis and Instrumentation (Xiamen University)
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An enthalpy-driven ligand is an ideal probe for practical applications because of the formation of abundant specific bonds between the ligand and target, compared to an entropy driven ligand with a similar Gibbs free energy change. However, there has been a lack of direct discovery strategy for identifying enthalpy-driven ligands. In this work, a molecular crowding SELEX (systematic evolution of ligands by exponential enrichment) strategy for discovering enthalpy-driven aptamers was developed to improve the affinity and selectivity of aptamers in complex samples. Three aptamer sequences were successfully evolved against a tumor biomarker Protein, and all proved to be enthalpy-driven by thermodynamics analysis, establishing the feasibility of molecular crowding SELEX for effective discovery of enthalpy-driven aptamers. Further comparison of aptamers evolved from conventional SELEX in buffer and molecular crowding SELEX (SYL-H2C) revealed much higher affinity of SYL-H2C. With its improved thermodynamic properties, the enthalpy-driven SYL-H2C aptamer was able to detect circulating tumor cells in real cancer patient blood samples with excellent detection accuracy (10/10). The proposed molecular crowding screening strategy offers a promising direction for discovering robust binding probes for a great variety of biomedical applications.
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