Table 2 Comparison of improved protein decoding methods

Flow cytometry

1.Nanoflow cytometry

2.form EV clusters,size larger than the detectable size of FCM

1.Increase detection limit

2.High stability, no blockage, no protein pollution

1.Equipment still needs to be developed

2.Complex equipment

Fluorescence imaging

1.Fluorescence spectrometry with

quantum dots

2.Branched rolling circle

amplification

3.Ring circle amplification digital detection

1.Plasma samples do not need pretreatment

2.High specificity, fast and cheap

3.Low detection limit and high accuracy

1.Limited accuracy

2.Limited capture type

3.Multi-step and long process

Thermophoresis Sensor

Thermophoresis aptamer sensor

Low cost and fast detection

and Simultaneous detection

of multiple marker proteins

Need better algorithms to eliminate the impact of pollution

SPR biosensor

1.Intensity-modulated, compact

SPR biosensor

2.nanoplasmonic EV

immunoassay utilizing

3.ECL immunosensor

1.Label-free, real-time and cost-effective detection

2.High sensitivity and

accuracy

3.Simultaneous detection of multiple EV surface proteins

1 Expense of sensitivity and accuracy

2.Only one protein can be detected at a time

3.Only surface protein can be detected

Integrated Magnetic-

Electrochemical EV(iMEX) Sensor

Integrated multi-channel separation and detection

1.Device portability

2.High throughput analysis

and detection

1.Poor versatility of equipment

Electrochemical sensor

1.iPEX (impedance

profiling of EV)

2.Anoparticle-Enabled Multiplexed electrochemical Immunoassay

3.Ultrasensitive electrochemical aptasensor

1.Rapid simultaneous

detection of multiple proteins

2.Simple detection with high sensitivity

3.High specificity, wide linear range

1.Only surface protein can be detected

2.Only surface protein can be detected

3.Complex materials