Bioscience seminar series
- Date: –09:30
- Location: Biomedicinskt centrum, BMC C4:2 Seminar room
- Lecturer: Prof. Masood Kamali-Moghaddam, Dept. of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University
- Contact person: Anna Nilsson
Molecular Tools for Parallel Detection and Characterisation of Proteins and Exosomes
Exosomes receive an increased attention in basic biology as well as in medicine. They are shown to be involved in many biological processes, and are proven to hold great potentials as diagnostic and therapeutic tools. However, there is an unmet need for new and improved technologies forquantitative and qualitative characterization of exosomes to meet challenges related to these vesicles, such as low concentrations in body fluids, the small size of the exosomes or the low copy numbers of antigens present on the surface of the exosomes.
We have developed a large number of affinity-based proximity assays for single- and multiplex detection of proteins and large complexes with high specificity and sensitivity. Several of these technologies, such as proximity ligation assay (PLA) combined with flow cytometry readout,
multiplex proximity extension assays (PEA) and proximity barcoding assays (PBA), are used for sensitive detection and characterization of individual exosomes.
Commonly, in these assays the exosomes are recognized by several affinity binders, each equipped with a DNA oligonucleotide. Upon binding of the target exosomes by the affinity probes, the DNA oligonucleotides are brought in proximity, subjected to enzymatic ligation or polymerization, which results in formation of an amplifiable reporting molecule. The use of
multiple recognition events in combination with signal amplification allows detection of exosomes with high specificity and sensitivity.
Here, I discuss the application of proximity assays for screening and validation of protein biomarkers, and for characterization and sensitive detection of exosomes in body fluids.