Abstract
Matrix metalloproteinase 2 (MMP2) plays critical roles in various diseases, such as atherosclerosis and cancer, and has been suggested to contribute to the instability of atherosclerotic plaque. To visualize MMP2 in pathologic tissues, we developed an aptamer targeting MMP2 protein by performing eight rounds of modified DNA systematic evolution of ligands by exponential enrichment (SELEX). The aptamer showed high affinity for MMP2 (Kd = 5.59 nM), precipitated MMP2, and detected MMP2 protein in pathological tissues such as atherosclerotic plaque and gastric cancer tissues. Furthermore, a MMP2 aptamer-conjugated fluorescent nanoprobe successfully visualized atherosclerotic plaques in apolipoprotein E (ApoE) knockout mice. These results suggest that the devised MMP2 aptamer could be useful for the development of various diagnostic tools.
Highlights
Matrix metalloproteinases (MMPs) are zinc- and calciumdependent proteolytic enzymes [1,2]
By performing modified DNA systematic evolution of ligands by exponential enrichment (SELEX), we successfully developed a Matrix metalloproteinase 2 (MMP2)-specific aptamer which had high affinity and specificity and showed the possibility that it can be applied for molecular imaging
MMP2 aptamer or anti-MMP2 antibody (AB37150, Abcam, Cambridge, England, UK) binding was performed at a dilution of 1:200 in blocking buffer overnight at 4°C, and secondary antibody binding was performed for 2 h at RT
Summary
Matrix metalloproteinases (MMPs) are zinc- and calciumdependent proteolytic enzymes [1,2]. Whereas many MMPs are secreted by cells, others are anchored on cellular membranes Members of this family play important roles in various cellular processes, such as migration, differentiation, and proliferation. Aptamers have several beneficial characteristics, such as low immunogenicity, low molecular weight (8 to 15 kDa), high stability, better penetration, high affinity, and ease of production [9]. From these reasons, we decided to develop a MMP2-specific aptamer. By performing modified DNA systematic evolution of ligands by exponential enrichment (SELEX), we successfully developed a MMP2-specific aptamer which had high affinity and specificity and showed the possibility that it can be applied for molecular imaging
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