Table 3 Lectin-based studies of EVs

PNA, MPA, EEA, MAL-I, MAL-II, AIA, STA

Tn antigen and α2,3-linked sialic acids

Healthy donor urine (uEVs)

IA and UC

Lectin- and antibody-captured uEVs show variations in size and surface glycans

Lectin microarray

[113]

WGA, ECL, AAL, PHA-E, WFA, PNA, ConA, SNA, MAL

T-antigen, N-glycan with bisecting GlcNAc and LacdiNAc structure, and α2,3-linked sialic acids

OVMz ovarian cancer cells

UC

Identified specific glycosignature and blocking of those glycosignature has impact on EV composition

Lectin blotting

[39]

ConA, SNA, MAL

LGALS3BP, complex N-glycans of the di-, tri-, and tetraantennary type with fucose, mannose, and bisecting GlcNAc

SKOV3 ovarian cancer cells

UC

EV internalization by recipient cells

Lectin analysis/blotting

[114]

Sialoglycoproteins & N-glycans found on the EV surface

Lectin blotting

[15]

PHA-M, ConA

Mannose, GlcNAc, GalNAc, and galactose

PCa cells and urine samples (uEVs)

Lectin agglutination

Lectin-induced agglutination of uEVs shows upregulation of PCa-associated mi-RNA

Lectin-immunoaffinity

[115]

WGA, LEL, STL, RCA, BPL, DSL, CAL, DBL, WFL

GlcNAc, and LacNAc oligomers

Healthy donor urine (uEVs)

IA and UC

lectin-based separation of uEVs shows a simple way of biomarker discovery

Lectin microarray

[51]

A panel of 47 lectins

N-glycans, and terminal sialic acids

Murine hepatic cell lines AML12 and MLP29

UC

Role of surface glycans in EV uptake

Lectin microarray

[116]

AIA, PNA, MPA, ABL, RCA-1

Core fucose and α2,3-linked sialic acids

Polycystic kidney disease (PKD) and healthy donor urine (uEVs)

UC and CF

uEVs glycoprofiling shows distinct patterns in PKD vs healthy individual

Lectin microarray

[117]

AAL, WFA, MAA, SNA

α2-3-linked sialic acids, and core fucosylated N- glycan

Cell lines of human HEK293 and glioma H4, and mouse glioma Tu-2449

UC and CF

structure elucidation and validation of N-glycan of EVs

Lectin blotting

[50]

Gal-1, DSL, ConA, AIA, GNA, NPA, PSA, UDA, HHL, CVN, SVN, GRFT, WGA

High-mannose, LacNAc, complex N-glycans, and fucose

H9, SupT1, Jurkat-Tat-CCR5 cells

UC

Glycomic profiling of HIV, cell membrane, and EVs derived from T-cells

Lectin microarray

[118]

A panel of 74 lectins

High mannose and complex N-linked glycans, polylactosamine, and α-2,6-sialic acids

SkMel-5, HT-29, HCT-15, H9, SupT1, Jurkat-Tat-CCR5, and breast milk

UC

Analyzing conserved glycan patterns of EVs

Lectin blotting and lectin microarray

[34]

PHA-M, ConA

Mannose, GlcNAc, and GalNAc

Blood, urine, ascites, and pleural liquids

UC and lectin agglutination

Lectin-aggregated EVs separation approach and characterized those EVs through proteomic studies

Lectin-induced agglutination, lectin blotting

[119]

GNA, DSA, PNA, SNA, MAA

Oligomannose

Astrocyte-enriched primary cultures (mouse)

UC

Synapsin is an oligomannose-binding lectin and releases from glial-derived EVs

ELISA, lectin staining

[120]

A panel of 45 lectins

α2-3 and α-2,6-linked sialic acids, fucose, and mannose

ADSC cells

UC

Glycan profiling of EVs and analysis of their uptake in vivo

Lectin microarray

[121, 122]

Specific glycan biomarker discovery for osteogenic differentiation

SSA

α-2,6-linked sialic acid

Melanoma cells

UC

Studied EV biogenesis and also identified membrane proteins on EVs as a membrane marker using a lectin

Lectin blotting

[123]

OAA

High mannose

Human glioblastoma, melanoma, colon and lung cancers

UC and lectin agglutination

A mannose binding lectin is used for affinity isolation of tumor derived EVs

Lectin-affinity approach

[30]

MAL-II, SNA, Jacalin, PNA, RCA120, GSA I-B4, DBA, SBA, Con A, sWGA, GSA-II, PSA, UEA-I, LTA

Galβ1, 3GalNAc, high mannose, and α-2,6-linked sialic acid

eAMCs

UC

Comparison of glycan composition on eAMCs and eAMCs-EVs

Lectin array

[124]

PHA-E, PHA-L, AAL

complex N-glycans with β1,6-branched tetraantennary and bisecting GlcNAc, and fucosylation linked (α1-3) or (α1-6)

MKN45 and glycoengineered MKN45 gastric cancer cell lines

UC, TEI, ODG, SEC

Different isolation approach yields distinct glycan associated EVs populations

Lectin blotting

[37]

SNA, ConA, Gal-3

Sialylated and mannosylated glycan structure

Seminal plasma from normozoopermic and oligozoospermic men

UC and gel filtration

Comparative analysis of EVs focusing their glycosylation

Lectin-TEM, and ion-exchange chromatography (IEC)

[41]

A panel of 45 lectins

Glycosylation of CD133 by sialic acids

Ascites samples and cell lines

TEI

Heavily glycosylated CD133 on EVs could be used as a potential biomarker for advanced pancreatic cancer

Lectin microarray

[18]

MAA, SNA, GNA, AAA, PHA-E and PHA-L

α-2,3 and α-2,6-sialic acids, fucoses, complex N-glycans with β1,6-branched tri/tetraantennary and bisecting GlcNAc

Primary WM115, WM793 and metastatic WM266-4, WM1205Lu melanoma cells

UC

EVs have specific glycan composition compared with their parental cell membrane fraction

Lectin-blotting

[53]

A panel of 48 lectins

High mannose, Galβ (1 → 3)-GalNAc, LacNAc, and GlcNAc

Adult helminth pathogen F. hepatica

UC

EVs released by parasite can interact with recipient host cells

Lectin microarray

[125]

SNA, Jacalin, PNA, HPA, UEA-I

Siaα-2,6Gal/GalNAc, fucose, T and Tn antigens

PANC-1 and HeLa cells

UC

Multiplexed detection of EV glycan pattern

Lectin array

[38]

A panel of 96 lectins, lectin rBC2LCN assay specific to EVs

Fucα1-2 Galβ1-3 GalNAc, Siaα-2,6 and LacNAc

hiPSCs and non-hiPSCs

Magnetic bead-based IA approach

Glycan profiles of EVs derived from stem cells

Lectin microarray and sandwich assay

[126]

A panel of 45 lectins

Galα1-3 GalNAc (T-antigen), and Siaα-2,3 Galα1-3 GalNAc (sialyl T-antigen)

Pancreatic cancer patient serum

Magnetic bead-based IA approach

Differential glycomic profiles of EVs and recognition of O-glycosylated EVs for pancreatic cancer detection

Lectin microarray

[35]

DBA, ECA, PHA-E4, MAM, WGA, ConA

GalNAc, GlcNAc, high-mannose, and Siaα2-3Gal

MDAMB231luc—D3H1 &-D3H2LN, and BMD2a breast cancer cells

UC

Identified altered glycosylation patterns on EVs which has inhibitory effect on EVs uptake

Lectin blotting

[127]

PHA-E

N-glycans with bisecting GlcNAc

MCF10A, MCF7, SKBR3, and MDAMB23 cell lines

UC

Modification of bisecting GlcNAc level on EVs has significant effect on metastasis of recipient cells

Lectin histochemistry

[128]

ConA, WGA, RCA, SNA, MAA, PHA-E, PHA-L, LCA, PNA, VVL, DBL

High-mannose, Siaα2-3Gal, and GlcNAcβ1.4

Human urine (uEVs)

UC

uEVs isolation from THP-depleted urine and use those uEVs for lectin-EV assay

Lectin-EV binding assay

[129]

AAL, SNA, PHA-E, PHA-L

Bisected and branched N-glycans, Fucα1-6GlcNAc, and α2,6-linked sialic acid

B16F10, Pan02, 4T1, AsPC1, MADAMB4175 cell lines

Asymmetric flow field-flow fraction (AF4)

Identification of EVs and their subpopulation

Lectin blotting

[43]

A panel of 34 lectins

Fucα1-2Gal, GalNAc, GlcNAc, mannose, and sialic acid

PCa cell line and bladder cancer patient urine (uEVs)

UC

Integrin-lectin assay significantly discriminate bladder cancer patients compared to benign control

Lectin-nanoparticle assay

[12]

SNA, ConA, AAL

α2,6-linked sialic acid, Fucα1-6GlcNAc, and mannose

Pancreatic cell line and patient serum

UC

Lectin conjugated janus nanoparticles (JNPs) specifically binds to glycans on pancreatic cell derived EVs and shows diagnostic potential

JNPs assisted- lectin-EV assay

[130]

A panel of 50 lectins

High-mannose, GlcNAc, GalNAc, and sialic acid

Helminth parasite F. hepatica

UC

Parasite EVs has influence on host-immune response during infection and glycosylated proteins of these EVs play role in EVs internalization

Lectin microarray

[42]