Multiplex detection and characterization of breast cancer exosomes by magneto-actuated immunoassay.

Multiplex detection and characterization of breast cancer exosomes by magneto-actuated immunoassay. Talanta. 2020 May 01;211:120657 Authors: Moura SL, Martín CG, Martí M, Pividori MI Abstract The exosomes are emerging as biomarkers for the detection of cancer in early stages, as well as for the follow-up of the patients under treatment. This paper describes the characterization of exosomes derived from three different breast cancer cell lines (MCF7, MDA-MB-231 and SKBR3), and the quantification based on a magneto-actuated immunoassay. The exosomes are separated and preconcentrated on magnetic particles by immunomagnetic separation and labelled with a second antibody conjugated with an enzyme for the optical readout performed with a standard microplate reader. Several molecular biomarkers, including the general tetraspanin CD9, CD63 and CD81, and the receptors related with cancer (CD24, CD44, CD54, CD326 and CD340) were studied either for the immunomagnetic separation or the labelling, in different formats. After a rational selection of the biomarkers, this immunoassay is able to detect 105 exosomes μL-1 directly in human serum without any treatment, such as ultracentrifugation. The interference from free receptors in the samples could easily be prevented by performing the immunomagnetic separation with antiCD81 modified magnetic particles and the labeling based on either CD24 or CD340. Furthermore, the differentiation of healthy d...
Source: Talanta - Category: Chemistry Authors: Tags: Talanta Source Type: research