Table 2. Details of Extracellular vesicle isolation and characterization techniques for their diagnostic role.

Study Name	Sample	Source of EVs	EV isolation technique	EV characterisation techniques used	EV concentration	EV size (mean, in nanometers)	Markers for EV validation
Ferrant J et al.1	SS and healthy controls	Plasma	Size-exclusion chromatography	NTA, flow cytometry	5×1010 (approx from graph)	110−150	NA
Aqrawi et al.2	SS and healthy controls	Tears and saliva	Size-exclusion chromatography	NTA, flow cytometry	Tears SS 1.09×109 ± 1.06×108 Control, 1.54×109 ± 3.08×108 Saliva SS 5.46×1010 ± 1.43×1010Controls 2.41×109 ± 3.98×109	Tears SS 171 ± 6.9Control, 163 ± 9.6Saliva SS 189 ± 4.1Controls 189 ± 4.4	CD9
Aqrawi et al.3	SS, non-SS and healthy controls	Tears and saliva	Size-exclusion chromatography	NTA, immunoaffinity capture, flow cytometry	Tears (SS, 5.0×108, Non-SS, 1.2×109Controls, 7.1×108) Saliva (SS, 1.9×1010± 0.7×109, Non-SS, 1.0×1010± 1.6×109 Controls, 7.9×109± 1.6×109	Tears (SS, 255 ± 40,Non-SS, 204 ± 8Controls, 215 ± 9) Saliva (SS, 233 ± 17,Non-SS, 231 ± 13Controls, 264 ± 6	CD9, CD−44
Kakan SS et al.4	NOD mice model	Plasma	Differential ultracentrifugation for discovery experiments and with SEC	RNA isolation, TEM, Western blotting, NTA	N/A	Median 136 nm (male NOD serum with UC), 131 nm (BALB/c mouse serum with UC).SEC-median diameter of 122 nm in both strains.	TSG101
Li F et al.5	SS and non-SS	MSGs and plasma	Total Exosome Isolation Kit (Invitrogen)	Not done	NA	NA	NA
Yamashiro K et al.6	SS and healthy controls	Mouth rinse	Ultracentrifugation	NTA, western blot	NA	145	CD9, CD63
Cross T et al.8	SS and healthy controls	Saliva	Size-exclusion chromatography and modified Qiagen exoRNeasy EV isolation	NTA, TEM, Immunoaffinity capture and Western blot	2.0 x109 particles/mL	154	CD9, CD63, Hsp70
Pucker et al.9	DED and non-DED	Tears	Polyethylene glycol (PEG) 8000	TEM, ELISA	550,000 in DED 600,000 in non DED	NA	CD63
Cross T et al.10	DED	Tears	Modified Qiagen exoRNeasy column approach	NTA, TEM, Flow cytometry, western blot	~108 particles/mL	100−350	Tetraspanin CD9 (flow cytometry), CD9, CD−63, Hsp70

SS − Sjögren’s Syndrome, DED − Dry Eye Disease, NTA − Nanoparticle Tracking Analysis, NOD − Non-Obese Diabetic, TEM − Transmission Electron Microscopy, SEC − Size Exclusion Chromatography, UC - Ultracentrifugation, MSG − Minor Salivary Gland