Selective Lipoprotein Removal Enables High‐Purity EV Isolation from Plasma via Aptamer‐Based Mesh Filtration

ABSTRACT Lipoproteins pose a major challenge to plasma extracellular vesicle (EV) isolation due to their abundance and physical similarity to EVs. Here, we present ApoFilter, a rapid and selective aptamer‐based mesh‐filtration platform that efficiently depletes ApoA1‐ and ApoB100‐positive lipoproteins. High‐affinity aptamers immobilized onto nylon mesh layers capture targeted lipoproteins in a spin‐column format, enabling rapid (1 min) separation under gravity‐driven flow. ApoFilter removed >99% of targeted lipoproteins (HDL and (V)LDL) from purified and plasma samples, with negligible capture of non‐target components and no compromise to EV integrity. When integrated with conventional EV isolation methods—including ultracentrifugation (UC), size‐exclusion chromatography (SEC), and ExoTFF—ApoFilter consistently eliminated lipoprotein contaminants across all workflows. This simple and rapid system addresses a key bottleneck in EV research, enabling the collection of ultrapure EVs suitable for downstream molecular analyses and translational applications.