Biology subjects

SHIN, S.

Publications and source records attributed to SHIN, S..

3 recordsLinked to original sources

bioRxiv2025

A Novel Aptamer-Based Approach for Lipoprotein Removal to Achieve Ultra-Pure Blood EV Isolation

Blood extracellular vesicles (EVs) are nanoscale lipid-bilayer particles that carry proteins, nucleic acids, and metabolites, rendering them powerful tools for non-invasive liquid biopsy and targeted drug delivery. However, clinical translation of blood-derived EVs is severely limited by the co-isolation of lipoproteins--whose size (30-100 nm) and density overlap with EVs--resulting in contaminated preparations that compromise biomarker accuracy and jeopardize therapeutic safety, efficacy, and biodistribution. To overcome this critical bottleneck, we developed ApoFilter, an aptamer-based affinity filtration platform engineered to selectively capture ApoB100- and ApoA1-containing lipoproteins (VLDL, LDL, HDL) while preserving EV integrity. In solutions containing only mixed lipoproteins, ApoFilter demonstrated a capture efficiency of 97% for ApoB100- and ApoA1-containing particles, yielding filtrates with >99% depletion of target lipoproteins. Crucially, this high selectivity was preserved in human plasma--a complex protein milieu--where ApoFilter removed over 99% of lipoproteins without any detectable loss of EV yield. Furthermore, when size-exclusion chromatography (SEC) or ExoTFF was integrated with ApoFilter, the limitations of each technique were innovatively complemented, achieving a highly advanced level of separation and purification. Notably, the ApoFilter-ExoTFF combination delivered the highest performance, attaining 99.9% protein removal and a 98.2% EV recovery rate, enabling ultrapure EV isolation from blood plasma. This synergistic purification strategy addresses both analytical and therapeutic requirements by eliminating lipoprotein interference in downstream molecular profiling and minimizing off-target effects in drug delivery. ApoFilter thus represents a versatile, scalable solution for isolating clinically relevant EVs from blood, substantially improving the reliability of liquid biopsy assays and accelerating the development of EV-based therapeutics.

bioengineering↗

bioRxiv2025

Scalable, high-throughput isolation of extracellular vesicles using electrokinetic-assisted mesh filtration

As extracellular vesicles (EVs) are increasingly recognized for their superior functions for therapeutics, the need for large-scale EV isolation technology is becoming more critical for clinical and industrial applications. Most existing EV isolation methods are optimized for small-scale laboratory samples, limiting their efficiency and scalability for large-scale production. Here, an electrokinetic-assisted filtration system (ExoFilter), which introduces charge interaction into physical mesh flow filtration, is proposed as a new candidate to address the challenges of scalable EV isolation. The hybrid filtration system demonstrates outstanding high-throughput EV isolation performance (a flux of [~]750 mL/min) using only a coarse physical filter, by electrokinetically arresting EVs flowing through the filter lattice. Furthermore, the recovery efficiency of ExoFilter, analyzed based on the ELISA results, was found to be approximately 98%, demonstrating the filters exceptional efficiency in EV isolation. Additionally, ExoFilter enables the rapid isolation of EVs from small samples as little as 200 {micro}L, facilitating quick and easy blood-based EV research. Furthermore, low-molecular-weight albumin from plasma samples was effectively removed. The high-throughput and high-efficiency characteristics of ExoFilter make it well-suited for scalable EV production, offering greater convenience for various clinical applications. HighlightsO_LIElectrokinetic-assisted mesh filtration (ExoFilter) enables scalable and rapid isolation of extracellular vesicles (EVs). C_LIO_LIA high throughput of [~]750 mL/min is demonstrated while maintaining high yield and purity. C_LIO_LIExoFilter effectively removes albumin contaminants from EVs through size-exclusive electrokinetic-assisted mesh filtration. C_LIO_LIEfficient EV isolation performance is achieved for human plasma, saliva, urine, and cell culture media. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=167 SRC="FIGDIR/small/645682v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@7ac3d5org.highwire.dtl.DTLVardef@1ced638org.highwire.dtl.DTLVardef@521c63org.highwire.dtl.DTLVardef@c5bd49_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

bioRxiv2025

Advanced Extracellular Vesicle Isolation: A Hybrid Electrokinetic-Tangential Flow Filtration Approach for Improved Yield, Purity, and Scalability

As extracellular vesicles (EVs) become increasingly important in diagnostics and therapeutics, achieving both high purity and yield during isolation remains a critical challenge. Conventional techniques often suffer from the co-isolation of non-vesicular particles and soluble proteins, limiting their clinical and research utility. In response, we introduce ExoTFF, a hybrid isolation technology that sequentially integrates electrokinetic filtration (ExoFilter) with size-exclusion tangential flow filtration (TFF) to deliver unprecedented performance gains through an iterative, synergistic mechanism. In the ExoTFF system, the sample is repeatedly circulated through an electrokinetic mesh filter and TFF until the liquid is removed. This recirculating flow gradually eliminates contaminants, while the electrokinetic filter continuously captures EVs as the sample is purified. Finally, any residual impurities in the TFF unit are completely removed via a dead volume elimination process. The complementary actions of these two distinct separation mechanisms double EV recovery rates and reduce impurity levels by 80% compared to conventional TFF, culminating in an impressive 800% improvement in the purity ratio. In proof-of-concept experiments, ExoTFF processed 10 mL of plasma within 10 minutes, efficiently depleting albumin and HDL while achieving superior EV recovery. To further explore scalability, an automated ExoTFF system processed 500 mL of sample in 50 minutes, maintaining consistent yield and purity. The ability to sustain performance across different scales highlights ExoTFFs potential for both laboratory research and industrial-level EV production. Beyond biological applications, this platform also offers broad applicability for the isolation of negatively charged nanoparticles, demonstrating its potential impact across multiple nanotechnology-driven fields.

bioengineering↗