A Dual-Signal Amplification Detection of Tumor mRNA Vaccines Based on the Novel Full-Length Catalytic Loop Initiating DNAzyme

The potential of mRNA vaccines, in the field of tumor treatment, has gradually emerged. With the development of mRNA vaccines, monitoring their levels has become particularly important. In this part of the work, a novel dual signal amplification strategy, based on the new "sensitive loop" activation full-length DNAzyme and DSN enzyme, was proposed, as an innovative method for detecting tumor mRNA vaccines. The novel full-length catalytic loop initiating DNAzyme (NFCL DNAzyme) is a single-stranded DNA molecule, consisting of a recognition loop (sensitive loop) and a catalytic core. This design abandoned the multicomponent pattern of traditional DNAzymes. When the target was present, it could partially bind to the capture probe. The partial capture probe, cleaved by DSN enzyme, could bind to the NFCL DNAzyme detection loop and activate the NFCL DNAzyme catalytic core to generate multiple amplifications. In addition, magnetic beads were introduced which could not only perform first-level signal amplification but also retain NFCL DNAzyme to fix the high cleavage activity sequence. This method broke through the limitations of traditional methods and greatly improved its universality. It aims to provide a new method for screening tumor mRNA vaccines and pharmacokinetic studies.