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Capillary electrophoresis-SELEX(CE-SELEX)
Introduction
The integration of capillary electrophoresis into SELEX was driven by the need for improved separation and analysis of nucleic acid sequences during the selection process. Mendonsa and Bowser first used this method in a work they published in 2004.
The key character of CE-SELEX is the use of CE for separating and analyzing nucleic acid sequences based on their size, charge, and conformation, allowing for high-resolution separation and detection of target-binding ligands.
Initial studies demonstrated the feasibility of using CE for the separation and analysis of nucleic acid pools enriched with target-binding ligands. One of the primary challenges in developing CE-SELEX was adapting the SELEX process to incorporate CE as a separation method without compromising the efficiency and robustness of the selection process. Optimizing CE conditions, such as buffer composition, voltage, and capillary dimensions, to achieve optimal separation of nucleic acid sequences was another difficulty.
The development of CE-SELEX was marked by the pioneering work of researchers who sought to integrate CE into the SELEX process. It has been successfully applied for the selection of aptamers against a variety of targets, including proteins, small molecules, and cells.
CE-SELEX offers several advantages over traditional SELEX methods, including higher resolution separation, shorter analysis time, and reduced sample consumption. The high-throughput and automation capabilities of CE-SELEX make it a valuable tool for the discovery of nucleic acid ligands with potential applications in therapeutics, diagnostics, and biotechnology.
Process
CE-SELEX retains the iterative process of SELEX, involving selection, partitioning, amplification, and enrichment of nucleic acid sequences. CE-SELEX involves the incubation of a nucleic acid pool with the target molecule, followed by separation of bound and unbound sequences using CE. The bound sequences are then eluted from the capillary, amplified, and subjected to subsequent rounds of selection to enrich for high-affinity ligands.