Many analytical techniques benefit greatly from the use of affinity reagent pairs, where each reagent recognizes a discrete binding site on the target. Nucleic acid-based aptamers provide many advantages over protein-based affinity reagents, including mature chemical synthesis, thermal stability, and low production costs. For the production of suitable aptamer pairs, Creative Biolabs has proposed a new method to effectively select aptamer pairs beyond the traditional systematic evolution of ligands by exponential enrichment (SELEX) technology.
Aptamer Pair
Compared with univalent receptor-ligand interactions, multivalent molecular systems usually produce higher affinity and specificity. In nature, microorganisms often use multivalent combination to gain advantages. Many molecular diagnosis and analysis techniques also use multivalency in the form of affinity reagent pairs, each of which binds to different sites of the target protein. For example, antibody pairs have been widely used to significantly improve the specificity of enzyme-linked immunosorbent assay (ELISA). These advantages have also been demonstrated for aptamers, where the use of aptamer pairs that bind to different sites of the target protein has shown significant performance improvement. However, the generation process of aptamers is a major technical challenge, and it is urgent to choose an effective and unbiased alternative strategy for aptamer pairs combined with different sites.
Methods of Aptamer Pair Development in Creative Biolabs
Traditional SELEX methods are based on target immobilization. In this method, candidate aptamers can access the target that does not occupy the target surface from the immobilization. Since aptamer pairs must bind to the same target at different sites, immobilization-free method has advantages in screening aptamer pairs. For example, graphene-oxide SELEX (GO-SELEX), target immobilization-free of SELEX, is a method that has successfully developed aptamer pairs. GO-SELEX is based on the strong adsorption of single-stranded DNA on the surface of graphene oxide by π-π stacking. Research reports that based on the use of GO-SELEX, aptamer pairs for one protein target and two viral targets have been successfully screened.
In addition, multiple aptamer isolation SELEX (MAI-SELEX) is also commonly used to screen aptamer pairs. MAI-SELEX has two different selection stages. The affinity module enriches the binding to a target on a single binding site, and the specificity module can group aptamer candidates based on the binding site.
Fig. 1 Overview of the multiplexed selection of aptamers SELEX process.1
What Can We Do?
The successful development of aptamer pairs has led to the development of sandwich biosensors for on-site diagnosis, similar to ELISA kits. Based on the application of SELEX technology platform, we develop new aptamer pairs for various targets through simple, low-cost and convenient aptamer screening methods, and customize each project to ensure that aptamers will play the best role in its final form. If you are interested in our service, please contact us for your exclusive solution.
Reference
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1.Yoshikawa, Alex M., et al. "A system for multiplexed selection of aptamers with exquisite specificity without counterselection." Proceedings of the National Academy of Sciences 119.12 (2022): e2119945119.
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Questions & Answer
A: The process typically includes target selection, aptamer generation, affinity maturation, characterization, and validation. Initially, target molecules are identified, and aptamers are generated using techniques like SELEX. The aptamers are then subjected to iterative rounds of optimization to improve their binding affinity and specificity. Finally, the aptamer pairs are characterized and validated for their performance.
A: Aptamer pairs developed through these services have a wide range of applications. They can be used in diagnostics for sensitive detection of disease biomarkers, in therapeutics for targeted drug delivery, and in biosensors for the detection of various analytes. Aptamer pairs also have potential applications in imaging, proteomics, and cell biology research.
A: Aptamer pairs offer several advantages over single aptamers. They can improve binding specificity by requiring simultaneous or sequential binding events, resulting in lower off-target interactions. Aptamer pairs can also enhance binding affinity, sensitivity, and selectivity. Additionally, the combination of two different aptamers in a pair can provide complementary recognition capabilities.
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