Journal: Journal of Hepatocellular Carcinoma

Article Title: A Single-Step Immunocapture Assay to Quantify HCC Exosomes Using the Highly Sensitive Fluorescence Nanoparticle-Tracking Analysis

doi: 10.2147/JHC.S423043

Figure Lengend Snippet: Overview of the experimental workflow of fluorescent-nanoparticle tracking analysis (F-NTA) to quantify HCC exosomes in their native state. ( A ). Shows antibody selection to immunocapture HCC exosomes. PE-labeled AFP and GPC3 antibodies are specific to HCC-derived exosomes used alone or in combination. ( B ). Brief presentation of clinical assay used to capture highly purified HCC exosomes for F-NTA quantification. 10μL of human serum was diluted in 100μL of PBS with 1% BSA. Antibody incubation was performed overnight at 4°C with shaking. The next day, the sample was diluted in 1mL of ultrapure-filtered distilled water and passed through the Sephadex column. Samples were further diluted 1:10 in ultrapure water and analyzed by NTA in light scattered mode (LSM) and fluorescence mode (FM).

Article Snippet: Sephadex G-200 (“hand-made gel column”, A 50120, Sigma, Burlington MA), Bio-Spin Chromatography Columns (7326008 Bio-Rad, Hercules, CA), Cell guidance Exo-spin mini columns (EX03-50), and Amersham MicroSpin G-50 Columns (27533001, Cytiva, Marlborough, MA) were used for this step.

Techniques: Selection, Labeling, Derivative Assay, Purification, Incubation, Fluorescence