A review of exosome isolation based on membrane protein recognition: Methods, performance, and challenges

Exosomes participate in physiological and pathological processes such as intercellular communication, immune regulation, and tumor initiation and progression, holding significant potential in early disease diagnosis, therapeutic target identification, and drug delivery system development. However, the isolation of exosomes presents significant challenges: their minute volume, high heterogeneity, extremely low yield, and complex biological matrix, among others. Traditional methods such as ultracentrifugation and ultrafiltration exhibit limitations including operational complexity, insufficient purity, and poor specificity. Researchers have increasingly focused on recognition capabilities of surface-specific membrane proteins on exosomes, developing separation strategies targeting these membrane proteins to achieve higher specificity and efficiency.

This review summarizes recent advances in exosome isolation strategies based on membrane protein recognition. The fundamental principles of affinity based capture are described, highlighting the selective interactions between exosomal surface proteins and recognition molecules such as antibodies, aptamers, and peptides. Representative technological platforms are further outlined, including immunomagnetic separation, solid phase affinity interfaces, microfluidic immunoaffinity systems, and ligand based enrichment approaches. These strategies demonstrate improved selectivity, capture efficiency, and analytical performance compared with conventional physical isolation methods. Recent developments integrating nanomaterials, microfluidics, and multifunctional platforms are highlighted for their ability to enhance analytical sensitivity and enable more precise exosome characterization. Current challenges, including exosome heterogeneity, limitations of recognition molecules, preservation of vesicle integrity, and the need for automated and high throughput systems, are also discussed, together with future perspectives for developing more robust, selective, and clinically applicable isolation technologies.

This article reviews the research progress of exosome isolation based on membrane proteins, discusses the advantages and challenges, and provides a reference for the optimization and application of exosome isolation methods in future work. The exosome isolation technology based on membrane proteins has higher specificity and efficiency than traditional physical methods. This article proposes future directions such as multi-marker identification, mild release, and automated clinical platforms to promote the clinical translation of exosomes.