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Table 2 New technologies for exosomal protein detection

From: Exosomes as a new frontier of cancer liquid biopsy

Methods Exosome sources Sample volume Sensing mechanism Sensing substances Detection limit Ref.
Colorimetric Detection MCF-7 cells and breast cancer patient’s serum   H2O2-mediated oxidation of TMB s-SWCNTs; CD63-specific aptamer 5.2 × 105 particles/μL [109]
Cell-culture medium and prostate cancer patient’s plasma 500 μL H2O2-mediated oxidation of TMB Aptamer-capped Fe3O4 nanoparticles 3.58 × 106 particles/mL [72]
Urine 100 mL H2O2-mediated oxidation of TMB Biotinylated anti-CD63 antibody; streptavidin-labeled HRP 35.0 AU/mL [76]
MCF-7 cells and cancer patient’s serum 100 μL H2O2-mediated oxidation of TMB CD9, CD63 antibody mixture; HRP-labeled secondary antibody 2.2 × 104 particles/μL [110]
BeWo cell   H2O2-mediated oxidation of TMB Au-NP; Fe2O3NC 103 exosomes/mL [148]
Fluorescence Detection Plasma specimens from NSCLC and OVCA patients 30 μL Chemifluorescence reagents EpCAM, IGF-1R or CA125 antibodies; AP-conjugated secondary antibody, and the DiFMUP substrate 0.281 pg/mL; 0.383 pg/mL [36]
SKOV3 cells and plasma of OVCA patients 10 μL The reaction of SβG with FDG Biotin conjugated detection antibodies and streptavidin conjugated SβG 21 exosomes/μL [149]
MCF-7 and MDA-MB-231 cell culture medium 1 mL Fluorescent carbocyanine dye (DiO) CD63 antibody functionalized microbead and DIO labelling   [150]
Cell culture supernatant and serum from pancreatic cancer patients   Fluorescent carbocyanine dye (DiO) CD63 antibody-functionalized EXOchip   [20]
MCF-7 cells and blood samples from cancer patients 100-300 μL Fluorescent second antibody Anti-EpCAM antibody and Alexafluor®647-conjugated secondary antibody   [59]
Cell-culture medium and plasma from breast cancer patients   Fluorescent second antibody CD63 antibody-coated magnetic beads; fluorescent dye-conjugated antibodies 107 particles/μL [151]
A549 cancer cell line and plasma samples of lung cancer patients 0.5 μL Fluorescent aptamer TMR-aptamer 500 particles/μL [152]
Serum samples   Fluorescent aptamer CD63 aptamer-modified magnetic beads; Cy3-labeled short sequence 1.0 × 105 particles/μL [126]
Cancer cell line and plasma samples 500 μL Fluorescent aptamer TPE-TAs/aptamer complexes; graphene oxide surface 3.43 × 105 particles/μL [99]
MDA-MB-231 cell-culture medium and plasma from breast cancer patients 80 μL Fluorescence quenching GPC-1 antibody coated magnetic beads; CD63 aptamer 6.56 × 104 particles/μL [84]
Cancer cell line and serum samples   Fluorescence quenching FAM-labeled aptamers; graphene oxide 1.6 × 105 particles/mL [113]
Cancer cell line and blood samples   Fluorescence quenching Anti-CD63-PE/MoS2–MWCNT 14.8 × 105 particles/mL [153]
Electrochemical Detection Ovarian cancer cell lines and plasma from patients with ovarian cancer 10 μL Integrated magneto-electrochemical sensor Immunomagnetic beads; HRP-labeled secondary antibody 3 × 104 [128]
Plasma samples 20 μL Electrochemical biosensor Immunomagnetic beads; probing antibodies   [129]
Cell-culture medium and blood samples from breast cancer patients   Electrochemical biosensor Anti-PD-L1-linked DNA strand; PVP@HRP@ZIF-8 334 particles/mL [114]
HepG2 cells and human serum of liver cancer patients 30 μL Aptamer-based biosensors CD63 aptamer and mimicking DNAzyme sequence 4.39 × 103 particles/mL [65]
Culture medium of HepG2 cells   Aptamer-based biosensors NTH-assisted aptasensor 2.09 × 104/mL [154]
Cell-culture medium and serum   Aptamer-based biosensors Aptamer-magnetic bead bioconjugates; electroactive Ru (NH3)63+ 70 particles/μL [155]
Cellular supernatant and human plasma samples   Aptamer-based biosensors anti-CD63 antibody modified gold electrode and a gastric cancer exosome specific aptamer 9.54 × 102/mL [156]
Human hepatoma cell lines MHCC97H/L and mouse melanoma cell lines B16-F1/10   Antibody microarray SPRi sensor Anti-CD9, CD41b,21 and tyrosine kinase receptor MET8a antibodies immobilized gold-coated glass sensor chip   [157]
SPR Detection MCF-7 breast cancer cells and MCF-10A normal breast cells   SPR-based aptasensor Dual gold nanoparticle-assisted signal amplification 5 × 103 exosomes/mL [158]
Human NSCLC cell lines, normal lung cell and plasma 1.5 mL Bioaffinity interactions of antibodies and different recognition sites Antibodies modified-gold chip and different recognition sites 104 particles/μL [159]
Urine samples from lung cancer patients and controls 500 μL SPR-induced improved scattering intensity Anti CD81/LRG1 antibody modified nanoporous gold nanocluster membrane; second antibody-conjugated Au nanorod probes < 1000 particles/mL [58]
Breast cancer cell line and serum 250 μL SPR-induced improved scattering intensity Anti-HER2-functionalised SPR chip 8280 exosomes/μL [160]
Cancer cells and serum and the CSF of an orthotopic mouse model   Strong localization of surface plasmon polaritons TIC-AFM and TiN–NH-LSPR biosensors 5.29 × 10−1 μg/ml; 3.46 × 10 -3 μg /ml [161]
Breast cancer cells and normal breast cells; plasma from HER2-positive breast cancer patients   Raman reporters Gold-coated glass microscopy slide; QSY21-coated gold nanorods 2 × 106/mL [134]
SERS Plasma of cancer patients 400 μL P-O bond signature Beehives-like Au-coated TiO2 macroporous inverse opal   [117]
Cell-culture medium and serum sample 4 μL MBA signature Fe3O4@TiO2 nanoparticles; anti-PD-L1 antibody modified Au@Ag@MBA 1 PD-L1 exosome/μL [90]
Normal and lung cancer cell lines; plasma   Deep learning Deep learning model   [162]
Cell-culture medium; serum and plasma < 1 μL Enrichment of aptamer-bound EVs Seven aptamers targeting specific proteins; machine-learning algorithm 3.3 × 103/μl [68]
CRISPR/Cas-assisted detection A549 cell-culture medium and serum from lung cancer patients   CRISPR/Cas12a CD63 aptamer; CRISPR/Cas12a Linear range of 3 × 103–6 × 107 particles/μL [136]
SUNE2 cell-culture medium and serum from NPC patients 50 μL CRISPR/Cas12a Nucleolin and PD-L1 aptamers; CRISPR/Cas12a 102 particles/μL [137]
SUNE2 cell-culture medium and serum from NPC patients   CRISPR/Cas12a CD109 and EGFR aptamers; CRISPR/Cas12a 102 particles/μL [138]
Single EV Analysis Human serum 10 μL Rolling circle amplification ssDNA-assisted single EV detection platform 82 vesicles/μL [139]
T3M4 pancreas cancer line and serum from PDAC patients 10 mL Flow cytometry Aldehyde/sulfate latex beads; anti-GPC-1 antibody and Alexa-488-tagged antibody   [38]
Breast cancer cell lines and serum of breast cancer patients 500 μL Flow cytometry Aldehyde/sulfate latex beads; anti-EpCAM or anti-HER2 antibody; Alexa-488- or − 594-tagged secondary antibodies   [141]
Human cell lines and serum of glioma patients 250 μL Flow cytometry Aldehyde/sulfate latex beads; anti-EGFR or anti-CXCR4 antibody   [118]
HCT15 cell-culture medium and plasma 500 μL Nano-flow cytometry    [142]
  1. OVCA Ovarian cancer, PGR Progesterone receptor, ESR1 Estrogen receptor 1, ERBB2 erb-b2 receptor tyrosine kinase 2