Matrix-Assisted Efficient Selection of a High-Affinity Circular Aptamer for Sensitive Detection of Low-Density Lipoprotein in Human Serum.

Abstract

The accurate and rapid detection of human serum biomakers is crucial for the early diagnosis of related diseases. Circular aptamers emerge as promising candidates for the precise recognition of these biomakers due to their remarkable biological and structural stability. In this study, we successfully identified a high-affinity circular DNA aptamer for Low-density lipoprotein (LDL), a key biomarker for cardiovascular diseases, through matrix-assisted efficient selection strategy conducted directly in human serum. This circular aptamer demonstrated a dissociation constant (Kd) as low as 18 nM and exhibited exceptional specificity for LDL, showing negligible responses to closely related analogs such as high-density lipoprotein and other abunzaidant human serum substances. We subsequently developed a sensitive aptasensor that utilizes the superior recognition capability of the circular DNA aptamer, which performed well even in human serum (with a limit of detection down to 0.4 μM and a wide linear concentration range of three orders of magnitude). This work underscores the feasibility of rapidly isolating highly functional circular DNA aptamers in one selection round, thereby accelerating the discovery of various circular DNA aptamers that are well-suited for molecular recognition of a wide array of emerging human serum biomarkers.