RESUMO
This review delves into the advancements in molecular recognition through enhanced SELEX (Systematic Evolution of Ligands by Exponential Enrichment) platforms and post-aptamer modifications. Aptamers, with their superior specificity and affinity compared to antibodies, are central to this discussion. Despite the advantages of the SELEX process-encompassing stages like ssDNA library preparation, incubation, separation, and PCR amplification-it faces challenges, such as nuclease susceptibility. To address these issues and propel aptamer technology forward, we examine next-generation SELEX platforms, including microfluidic-based SELEX, capillary electrophoresis SELEX, cell-based aptamer selection, counter-SELEX, in vivo SELEX, and high-throughput sequencing SELEX, highlighting their respective merits and innovations. Furthermore, this article underscores the significance of post-aptamer modifications, particularly chemical strategies that enhance aptamer stability, reduce renal filtration, and expand their target range, thereby broadening their utility in diagnostics, therapeutics, and nanotechnology. By synthesizing these advanced SELEX platforms and modifications, this review illuminates the dynamic progress in aptamer research and outlines the ongoing efforts to surmount existing challenges and enhance their clinical applicability, charting a path for future breakthroughs in this evolving field.
RESUMO
Aim: Investigating molecularly imprinted polymers (MIPs) in electrochemical biosensors for thrombin detection, an essential protein biomarker. Comparing different monomers to showcase distinct sensitivity, specificity and stability advantages. Materials & methods: Dopamine, thionine and ethanolamine serve as monomers for MIP synthesis. Electrochemical methods and atomic force microscopy characterize sensor surfaces. Performance is evaluated, emphasizing monomer-specific electrochemical responses. Results: Monomer-specific electrochemical responses highlight dopamine's superior signal change and stability over 30 days. Notably, a low 5 pg/ml limit of detection, a broad linear range (5-200 pg/ml) and enhanced selectivity against interferents are observed. Conclusion: Dopamine-based MIPs show promise for high-performance electrochemical thrombin biosensors, suggesting significant applications in clinical diagnostics.
Assuntos
Técnicas Biossensoriais , Impressão Molecular , Dopamina , Trombina , Polímeros/química , Impressão Molecular/métodos , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Limite de Detecção , EletrodosRESUMO
This study developed an aptamer-based enzyme-linked immunosorbent assay (ELISA) system for sensitive detection of the d -dimer molecule, which is significantly elevated in several severe diseases. Aptamers are known to have advantages, such as longer stability and usability at different temperatures and in different complex samples, over antibodies, which are generally used in traditional ELISA methods. In the present work, the aptamer-based ELISA system was capable of quantitatively determining in a wide range (100 ng/mlâ10 µg/ml) and remain stable for a couple of months even under ordinary room conditions. Validation of the developed system was evaluated by spiked human serum samples with high accuracy. With more comprehensive project steps, the developed aptamer-based ELISA system could be transformed into a platform that offers a sensitive, portable, and easy-to-use tool for analyzing d -dimer molecules.