HansaBioMed is your partner in accelerating discoveries in exosome research
The ideal platform to detect protein and RNA in exosomes
ExoTESTTM
Exosomes immunocapturing
Simple ELISA assay for exosomes capture, quantification and characterization from culture supernatants, plasma or other biofluids
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 Physiological exosome functions
Pathological exosome functions
Exosomes have pleiotropic functions dependent on cell type and condition
Emerging role in diverse pathological conditions such as cancer, infectious and neurodegenerative diseases
Export pathway for proteins that are to be discarded or to be passed on to other cells
Due to alteration in cell metabolism and export pathways the concentration of exosomes is elevated in the blood of cancer patients1; exosomal export of anti cancer drugs contributes to drug resistance2
Cell-cell interaction/signaling
Cancer spread and immunomodulation
Inter-cellular communication devices for transfer of proteins, lipids and genetic material between cells, in the parent cell microenviroment and/or at distance
Cancer spread - mRNAs and miRNAs present in exosomes can transfer new properties such as metastatic ability
Antigen presentation and immune response Immunomodulation – exsosomes are so far diagnostic purposes. Exosomes have been isolated from regulation (stimulation or suppression) implicated both in tumor rejection and inhibition of diverse cell lines (hematopoietic cells, tumor lines, antitumor immune responses primary cultures, virus infected cells) as well as from Formation of morphogen gradients during Transfer of infectious cargo (prions, viruses, β development amyloid 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). During their biogenesis exosomes incorporate a wide range of membranes and cytosolic proteins involved in many cellular functions 1-4 . In this way besides proteins that reflect their endosomal origin, exosomes contain a unique protein fingerprint that reflects parent cell type and condition. Exosomes shuttle also RNA molecules, mRNAs and miRNAs, both classes 5,6 proved to be functional in target cell, being translated or modulating gene silencing respectively . Consequently exosomes may act as inter-cellular communication devices that extend new properties to recipient cells. Overall, exosome molecular composition distinguishes resting from activated, healthy from diseased cell.
EXOTESTTM IS A FUNDAMENTAL TOOL FOR EXOSOME-BASED RESEARCH TM
ExoTEST is a patented double sandwich ELISA assay for quantitative and qualitative analysis of exosomes. TM ExoTEST captures exosomes from biological samples by using antibodies for exosome specific surface proteins. Quantification and characterization of exosomal proteins is subsequently performed by using detection antibodies of TM interest. By employing different combinations of capture and detection antibodies ExoTEST can be customized for assessing multiple antigens in a total exosome population in the sample or addressing a defined exosome subpopulation. Informative protein profile for the samples of interest can be combined/complemented with subsequent RNA (mRNA or miRNA) extraction and analysis from captured exosomes. The upstream enrichment in specific exosome subpopulation(s) significantly reduces the background noise that impairs the analysis of RNA sequences. TM ExoTEST therefore represent a fundamental novel, reliable, practical and easy solution for any comprehensive exosome analysis from different biological samples including human body fluids.
ExoTESTTM
Exosomes immunocapturing
EXOTESTTM FEATURES AND ADVANTAGES 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.
Scheme of the basic ExoTEST procedure 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 some immediate 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. 2. 3.
Exosome capture and quantification Exosome comprehensive profiling Specific applications (in research and diagnosis)
ExoTEST
tm
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
ExoTESTTM is a fundamental tool for any exosome-based research
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. This feature is currently only available as a service by HansaBioMed ( info@hansabiomed.eu).
Starting autumn ExoTEST will be also available for sale in special kits for exosome sub-populations selective capturing.
HIGH SPECIFICITY AND SENSITIVITY ExoTEST enables reliable and precise quantitative measurement and comparison among samples and individual experiments. Standard exosome preparations are provided for making standard curves and assay calibration. ExoTEST ensures dose dependent detection of exosomal markers from different biological samples including human biofluids such as plasma and urine. Assay is analytically validated and provides increased sensitivity in detection of exosomal markers with respect to other analytical methods. Example of standard curve obtained with exosomes derived from human HBM-Colo1 cell line (A) and titration of exosomes purified from human plasma (B) and urine (C).
ExoTEST enables both capture and analysis of exosomes purified from cell supernatants and human biofluids as well as directly from unfractioned samples.
ExoTESTTM is a fundamental tool for any exosome-based research
Example of quantification of exosomes from ultracentrifuged and unfractioned cell supernatant and human plasma samples (Logozzi 2009)
RESOLUTION BETWEEN DIFFERENT CELL TYPES AND BETWEEN HEALTHY AND DISEASED SAMPLES ExoTEST enables detection of cell type or disease specific antigens on exosomes captured from a sample. It is suitable therefore for the analysis of exosomes from human body fluids (plasma, urine) and detection of known and novel biomarkers in routine clinical samples, being a perfect tool for exosomal markers research and discovery. ExoTEST allows quantitative assessment specific exosomal populations defined by antigens differentially expressed under normal and pathological conditions. Elevated concentration of circulating exosomes carrying specific markers is reported in diseases such as cancer, inflammatory and infective diseases.
Example of quantification of specific exosomes displaying tumor specific markers in human plasma samples (Logozzi 2009)
COMPREHENSIVE EXOSOME PROFILING - MULTIPLE ANTIGENS AND RNAS By employing different combinations of capture and detection antibodies ExoTEST can be customized for assessing multiple antigens in a total exosome population in the sample or in a defined exosome sub-population. Characterization of samples of interest can be complemented with subsequent RNA (mRNA or miRNA) extraction and analysis from captured exosomes. The upstream enrichment in specific exosome sub-population(s) significantly reduces the background noise that impairs the analysis of RNA sequences. Combining proteic and RNA markers analysis for a single sample can provide highly informative marker profiles that further increase a resolution potential of the ExoTEST based platform.
ExoTESTTM is a fundamental tool for any exosome-based research
ExoTEST
TM
: The ideal platform to detect protein and RNA in exosomes
Products available starting July 2011: HansaBioMed is announcing its first products for exosome research which will be ready for shipping in July 2011. ExoTEST Basic kit is meant as an exosomes capturing tool, while the ExoTEST Ready to use kit is meant as an exosomes capturing and quantitative detection tool. Each kit include operative manual and positive controls. The first ExoTEST products available on the market starting July 2011 are the following: Price: 1. 2. 3. 4.
ExoTEST ELISA precoated plates for exosome capturing ExoTEST Basic Kit ExoTEST Ready to use Kit ExoTEST lyophilized standards starting from
€ 300,00 € 700,00 € 1300,00 € 250,00
Promotion price: (orders before 30.06.2011) € 225,00 € 525,00 SAVE € 975,00 € 185,00 MONEY
To have constantly updated information on our products and to make your orders please visit our website at www.exotest.eu/products or write us at orders@exotest.eu.
ExoTEST
TM
: The ideal platform to detect protein and RNA in exosomes
Custom services: In order to facilitate your exosome research HansaBioMed offers a wide range of custom services, allowing you to spend less time making your own tools and concentrate your efforts in making discoveries. Our services include: 1. Purification of overall exosomes from any sample of interest (cell supernatants, any animal or human biofluids) 2. Preparation of samples suitable for downstream protein and RNA profiling and biomarker discovery 3. Quantification and characterization of exosomal proteins by WB, FACS and ELISA 4. Comparative analysis of multiple samples for markers of interest 5. Preparation of calibration standards and quality controls from selected model cell lines suitable for long term storage We also provide scientific consultancy and assistance in interpretation of results. To have constantly updated information on our services and to obtain on offer for the service of interest please visit our website at www.exotest.eu/services or write us at info@exotest.eu.
ExoTEST
TM
: The ideal platform to detect protein and RNA in exosomes
Coming soon: HansaBioMed is working to release in the near future ExoTESTTM special kits for different applications in research use in the fields cancer and neurodegeneration and to provide you a wide panel of characterizing antibodies for research use, tested and certified for the ExoTESTTM platform. Moreover, HansaBioMed is working to validate a number of specific ELISA diagnostic kits, based on the ExoTESTTM platform, for clinical use in the following therapeutic areas: cancer and neurodegeneration.
HansaBioMed OU Akadeemia tee 15 12618 Tallinn ESTONIA Reg. No. 11393103 Vat No. EE101155126
Phone: +372 6204348 Mobile: +39 3453065144 Fax: +372 6204358 info@exotest.eu www.exotest.eu
©2011 HansaBioMed OU. All rights reserved. HansaBioMed, HansaBioMed logo and ExoTEST are trademarks of HansaBioMed OU.