IRTIDP: A simple integrated real-time isolation and detection platform for small extracellular vesicles Glypican-1 in pancreatic cancer patients.

Glypican-1 (GPC-1) protein-positive small extracellular vesicles (GPC-1+-sEV) have been proposed as potential biomarkers for early diagnosis of pancreatic cancer. In this study, we present an integrated real-time isolation and detection platform (IRTIDP) to capture and analyze GPC-1+-sEV directly from sera of pancreatic cancer patients. First, CD63 antibody-modified metal-organic framework (MOF) materials were utilized to enrich sEVs with a capture efficiency of 93.93 %. Second, a SERS probe was constructed by Raman reporter 4-MBA and GPC-1 antibody modified SERS active silver nanoparticles (AgNPs), which formed a sandwich complex structure of "MOFs@GPC-1+-sEV@AgNPs-4-MBA" with MOFs-enriched sEVs. The IRTSDP can complete the capture and detection process within 35 min, with a detection limit for 1 GPC-1+-sEV/μL, and linear range between 105∼109 GPC-1+-sEV/mL. Furthermore, this approach has been applied to quantify serum sEV GPC-1 in clinical pancreatic cancer patients. Based on the SERS intensity analysis, pancreatic cancer patients can be distinguished from pancreatic cystadenoma patients and healthy individuals effectively using this innovative platform that provides highly specific and sensitive means for early diagnosis of pancreatic cancer as well as other tumor types. [Display omitted] • The MOFs@CD63 capture probe can separate small extracellular vesicles (sEVs) in 10 min with a capture efficiency of 93.93 %. • The anti-GPC-1 antibody modified AgNPs@4-MBA was designed as a SERS nanotag for sEV GPC-1 labeling and accurate quantification. • By integrating MOFs@CD63 isolation and SERS immunoassay detection, a detection limit of 1 GPC-1+-sEV/μL can be achieved within 35 min. • Based on SERS analysis, pancreatic cancer patients can be distinguished from the pancreatic cystadenoma patients and healthy individuals by 50 μL serum samples. [ABSTRACT FROM AUTHOR]