Journal: bioRxiv

Article Title: Combining Anion Exchange and Size Exclusion Chromatography for Extracellular Vesicle Enrichment from Small Volumes of Human and Mouse Plasma for Quantitative Proteomics

doi: 10.64898/2026.03.11.711200

Figure Lengend Snippet: Workflow for EV enrichment from 4 ml (A) and 0.5 ml (B). EV markers (CD63 and Syntenin-1) and plasma contaminant proteins (Albumin and ApoA1(shown in red)) by western blot analysis (panel C). Starting plasma volume, final volume post concentration with the centrifugal filter and gel loading volume were used to calculate the concentration factor and equivalent plasma starting amounts indicated above figure.

Article Snippet: Membranes were blocked in 5% skim milk in PBS at room temperature for 1 hour before overnight incubation with primary antibodies Syntenin (Abcam, cat# ab133267), CD63 (Abcam, cat#ab134045), CD63 (Abcam, cat#ab8219), Albumin (Cell Signalling Technology, cat#4929S), Calnexin (Cell Signalling Technology, cat#2679S) ApoA1 (Santa Cruz, cat# SC-376818).

Techniques: Clinical Proteomics, Western Blot, Concentration Assay

Journal: bioRxiv

Article Title: Combining Anion Exchange and Size Exclusion Chromatography for Extracellular Vesicle Enrichment from Small Volumes of Human and Mouse Plasma for Quantitative Proteomics

doi: 10.64898/2026.03.11.711200

Figure Lengend Snippet: SEC Void optimisation. Pooled SEC fractions 1-4 collected using a 2.7 mL void volume (1-4A) and a 2.8 mL void volume (1-4B) were prepared in triplicate from 0.5 mL plasma. Western blot analysis of contaminant proteins Albumin (green) and lipoprotein ApoA1 (red) (A) with 0.5 µl plasma included as a positive control (+). EV markers CD63 and syntenin-1 and small EV exclusion marker Calnexin (B). Samples (0.5 ml and 0.4 ml) from Experiment 3.2 included as a western blot control and 2µl platelet lysate included as a calnexin positive control (+). Electron microscopy (C) visualisation and size and concentration histogram (D) of EV representative SEC sample (Void 2.7 ml).

Article Snippet: Membranes were blocked in 5% skim milk in PBS at room temperature for 1 hour before overnight incubation with primary antibodies Syntenin (Abcam, cat# ab133267), CD63 (Abcam, cat#ab134045), CD63 (Abcam, cat#ab8219), Albumin (Cell Signalling Technology, cat#4929S), Calnexin (Cell Signalling Technology, cat#2679S) ApoA1 (Santa Cruz, cat# SC-376818).

Techniques: Clinical Proteomics, Western Blot, Positive Control, Marker, Control, Electron Microscopy, Concentration Assay