Signal amplification strategies in biosensing of extracellular vesicles (EVs).

Extracellular vesicles (EVs) are membrane-enclosed vesicles secreted from mammalian cells. EVs act as multicomponent delivery vehicles to carry a wide variety of biological molecular information and participate in intercellular communications. Since elevated levels of EVs are associated with some pathological states such as inflammatory diseases and cancers, probing circulating EVs holds a great potential for early diagnostics. To this end, several detection methods have been developed in which biosensors have attracted great attentions in identification of EVs due to their simple instrumentation, versatile design and portability for point-of-care applications. The concentrations of EVs in bodily fluids are extremely low (i.e. 1-100 per µl) at early stages of a disease, which necessitates the use of signal amplification strategies for EVs detection. In this way, this review presents and discusses various amplification strategies for EVs biosensors based on detection modalities including surface plasmon resonance (SPR), calorimetry, fluorescence, electrochemical and electrochemiluminescence (ECL). In addition, microfluidic systems employed for signal amplification are reviewed and discussed in terms of their design and integration with the detection methods.

A microfluidic electrochemical aptasensor for highly sensitive and selective detection of A549 cells as integrin α6ß4-containing cell model via IDA aptamers.

There is a growing interest in developing microfluidic biosensors for the accurate and reproducible analysis of various biomarkers obtained from liquid biopsy. This paper reports a novel microfluidic electrochemical aptasensor for determination of A549 cells as integrin α6ß4-containing cell model and circulating tumor cell (CTC) model of NSCLC, based on target-induced structure switching mode. The conformational change of IDA aptamer structure with an affinity towards A549 cells, in the absence and presence of A549 cells allowed selective and sensitive detection of A549 cells. The microfluidic biosensor consisted of a microchip integrated with a screen-printed gold electrode functionalized with SH-IDA aptamers via covalent chemistry. Target solution containing A549 cells in Phosphate-buffered saline could be introduced to the microchip for analysis. Upon exposing to redox probe, a reduction in peak current occurred. Required flow sequencing for various steps of the detection protocol was performed using on-chip gas-actuated microvalves with programmable operation. The microfluidic biosensor provided a wide linear dynamic range of 50-5 × 105 cells/mL and allowed determination of A549 cells with a detection limit of 14 cells/mL. Furthermore, the microfluidic biosensor was efficiently used for detection of A549 cells in complex matrices such as human serum. Our novel microfluidic aptasensor presents an enabling analytical platform to perform various detections of low volume biomarkers with high accuracy and reproducibility.