Table 1. Overview of Glycosylation in EV Analysis Methods^a.

method	principle	advantages	disadvantages	ref
GC LC	physical properties of analyzed samples	suitable for analysis of a single glycoprotein	require standards to decode the glycosylation structures	(66−68)
MALDI-MS	identification of the glycan structures and glycosylation sites by m/z of fragments	simple and fast, only a few seconds, high sensitivity	incompatible with chromatography, only useful for glycosylation analysis in simple samples	(73−75)
ESI-MS		compatible with multiple chromatography, suitable for glycosylation analysis in complex samples	time-consuming, low throughput, high cost, complex spectra, difficult to decode	(78, 79, 92)
WB	transfer glycoproteins to membranes and detect with antiglycan antibodies	qualitative and semiquantitative analysis, validation analysis of glycoproteins	poor accuracy and reproducibility in quantification, antibody-dependent	(80, 82)
ELISA	antigen–antibody binding, quantification by fluorescence or luminescence signals	mature technology with good reproducible results, suitability for detecting extracellular secreted proteins	quantitative analysis of glycoproteins and glycans with known glycans structures	(83−85)
lectin-based method	lectins bind and recognize the carbohydrates structures	high sensitivity, lectin arrays with high throughput	low accuracy, disturbed and inhibited by other monosaccharide structures	(86,87,89,93)
fluorescence detection	detection by conjugates emission	fast, high resolution	requires fluorescent labeling prior to detection	(90, 94)

^aThe abbreviations in the table are explained with the corresponding full names as follows: GC, gas chromatography; LC, liquid chromatography; MALDI-MS, matrix-assisted laser desorption/ionizationmass spectrometry; ESI-MS, electrospray ionization mass spectrometry; WB, Western blotting; ELISA, enzyme-linked immunosorbent assay; and ref, references.