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Exosomes are small endosome-derived lipid nanoparticles (50-120 nm) actively secreted by exocytosis by most living cells. Exosome relase occurs either constitutively or upon induction, under both normal and pathological conditions, in a dynamic, regulated and functionally relevant manner so that both amount and molecular composition of released exosomes depend on the state of a parent cell. This is one of major premises of employing exosomes for research and diagnostic purposes. Exosomes have been isolated from diverse cell lines (hematopoietic cells, tumor lines, primary cultures, virus infected cells) as well as from biological fluids in particular blood (e.g. serum and plasma from cancer patients) and other body fluids (bronchoalveolar lavage fluid, pleural effusions, synovial fluid, urine, amniotic fluid, semen, saliva etc). Exosomes have pleiotropic physiological and pathological functions and an emerging role in diverse pathological conditions such as cancer, infectious and neurodegenerative diseases.


 

- Exosomes incorporate a wide range of membranes and cytosolic proteins involved in many cellular functions and reflecting both their endosomal origin and a protein fingerprint unique for parent cell type and condition.


- Exosomes shuttle also RNA molecules, mRNAs and miRNAs, both classes proved to be functional in target cell, being translated or modulating gene silencing respectively. Overall, exosome molecular composition distinguishes resting from activated, healthy from diseased cell.
ExoTEST is a novel, reliable, versatile and easy to use double sandwich ELISA assay for quantitative and qualitative analysis of exosomes.


ExoTEST platform consists ELISA plates pre-coated with proprietary pan-exosome antibodies enabling specific capture of exosomes from different biological samples, including cell culture supernatants and human biological fluids. Quantification and characterization of exosomal proteins is subsequently performed by using appropriate detection antibodies against exosome associated antigens that can be either common for all exosomes or cell type- or cell condition specific. ExoTEST enables detection of multiple exosome associated antigens indicative of type and condition of the parent cell, providing thus a platform for assessment of cell state and function.


By employing different combinations of capture and detection antibodies ExoTEST can be customized for assessing multiple antigens in a total exosome population as well as enrichment with cell/tissue specific exosomes from body fluids. Assay provides someimmediate readouts, namely origin, quantity and molecular composition of  isolated exosomes. For the samples of interest RNA (mRNA or miRNA) can be extracted and analysed from captured exosomes.

Therefore ExoTEST platform allows:

  1. Exosome capture and quantification
  2. Exosome comprehensive profiling
  3. Specific applications (in research and diagnosis) 

 

 

EXOTEST CHARACTERISTICS AND ADVANTAGES
ExoTEST kits consist in single well ELISA plates pre-coated with proprietary pan-exosome capturing antibodies, exosomes standards obtained from model cell cultures for assay positive control and/or plate calibration and a number of different primary antibodies for exosome detection and quantification.

ExoTEST main features:

  • 12x8 pre-coated plates are stabilized and suitable for long term storage
  • Available either transparent or white pre-coated plates enabling both chromogenic and luminometric detection
  • Flexible and multiplex assay design: a single ELISA plate allowes simultaneous assessment of multiple antigens in a single sample as well as screening of a large number of samples
  • Growing panel of detection antibodies that can be provided by HBM (or selected authonomously by the user)
  • Lyophilized and quantified exosome standards for assay calibration suitable for long term storage
  • It allows exosome capture and analysis from different biological samples (cell supernatants, plasma, urine, CSF)
  • It is a unique assay that enables contemporary capture, quantitative and qualitative assessment of exosomes from the sample
  • Simple and fast procedure

ExoTEST also permits exosome enrichment procedures (i.e. cell/tissue specific exosomes from biological fluids) in order to obtain homogeneous and well defined samples, allowing to select specific sub-populations of exosomes. 
Starting autumn ExoTEST will be also available for sale in special kits for exosome sub-populations selective capturing.