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1 Summary Section 7 Introduction 68 Exosome and Microvesicle Isolation 69 EV protein quantification, sceening and profiling 69 Nanoparticle Tracking Analysis (NTA) 70 EV Nucleic Acid isolation 70 Nucleic Acid profiling and NGS 71 Custom Products 72 Additional services and project collaborations 72
2 Customer support For information and customer support contact us at: Advantages Introduction Being the oldest company entirely dedicated to exosome research, since 2007 HansaBioMed Life Sciences has developed unique competences and tools allowing us to help you with your research work. Our experienced scientists will help you accelerate your research and achieve your objectives quickly, providing consultancy and/or professional services using state of the art equipment. Our services include: EV purification and services performed by experienced scientists and qualified personnel. Material is processed quickly. HansaBioMed Life Sciences has all the necessary facilities for exosome purification, protein and nucleic acid profiling. 68
3 Catalog 2018 Section 7 Exosome and Microvesicle Isolation HansaBioMed Life Sciences provides high expertise in Exosome and Microvesicle purification from human or animal biofluids and cell culture media (HBM-LS can provide both cell growth and EV isolation or EV isolation from supernatant collected by customer). Exosome isolation can be obtained following our standard protocols which combine tangential flow filtration and SEC, or via other methodologies, such as immunoaffinity (latex beads) and ultracentrifugation. Isolated EVs are subsequently quantified for total protein contents and validated by nano tracking analysis (NTA) with the Zetaview equipment (Particle Metrix). Purified EVs can be shipped to the customer lyophilized or frozen, as required. Cell culture growth and EV isolation Isolation by Ultracentrifugation and Microfiltration Isolation by Size Exclusion Chromatografy Isolation by Immunoaffinity Exosome and Microvesicle Isolation Cell lines can be shipped to HBM, who provides cell growth and EV isolation. Method for isolating EV from a large amount of sample. includes EV isolation, NTA and protein content quantification. Isoaltion from small volume (100 μl - 2 ml). includes EV isolation, NTA and protein content quantification. Isoaltion from small volume (500 μl - 10 ml). The method allows to enrich EV subpopulations. includes EV isolation, NTA and protein content quantification. EV protein quantification, screening and profiling Together with RNA, proteins are an important source of biomarkers and potential indicators of disease states. HansaBioMed provides services for detection, quantification and profiling of exosome proteins, which can be analyzed via ExoTEST ELISA, Mass spectrometry, FACS analysis or Western Blotting. EV protein quantification, screening and profiling ELISA assay is an efficient method for isolating and profiling protein markers. HBM-LS can provide: 1- Multiple profiling of exosome markers from a single sample. EV ELISA profiling 2- Exosome marker profiling of a large number of samples. 3- Possibility to customize the ELISA plate for EV subpopulation enrichment. FACS analysis on pre-isolated EVs and EV isolation and FACS FACS analysis of EV markers analysis. Mass Spectrometry analysis of Mass Spectrometry profiling on pre-isolated EVs. EV markes Western Blotting analysis Western blotting analysis and profiling of EV markers. 69
4 Nanoparticle Tracking Analysis service Nanoparticle tracking analysis (NTA) is a method for visualizing and analyzing particles in liquids that relates the rate of Brownian motion to particle size. NTA allows the determination of a size distribution profile of small particles with a diameter of approximately nanometers (nm) in liquid suspension. NTA service is performed by the use of Zetaview (Particle Metrix), which allows EV characterization by size distribution, concentration, zeta potential and phenotyping. EV Nucleic Acid Isolation EVs shuttle functional RNAs (75% mirna) and genomic DNA. HansaBioMed Life Sciences provides a wide portfolio of kits and services for EV-associated RNAs and DNA isolation from human/ animal biofluids (plasma, serum, urine, etc.) and cell culture media. Nucleic Acids (NA) are isolated with the HBM-LS proprietary technology and quantified by Nanodrop analysis. Delivery time approximately 1-2 weeks. EV-RNAs Isolation EV-DNA Isolation EV Nucleic Acid Isolation EV-RNA is purified by HBM-LS RNA extraction kits, which uses immunobeads for capturing EVs. Customers have the advantage to customize the immunobeads for enrichment of specific EV subpopulations 1-Total RNA from overall EV population 2-Total RNA from specific EV subpopulations DNA isolation is performed by HBM-LS kit. The service allows: 1- Isolation of cf-dna and EV-DNA 2- Isolation of only EV-DNA 3- Isolation of genomic DNA contained in EV subpopulations. EV Nucleic Acid profiling and Next Generation Sequencing Hansabiomed Life Sciences scientists can provide an expert service for the identification of exosome-associated RNA/DNA biomarkers leveraging the throughput and scalability of Ion Torrent and SOLiD System (Thermo Fisher Scientific) next-generation sequencing (NGS) platforms. This service is an easy solution for researchers who are interested in identifying new exosome RNA biomarkers or in the profiling expression of such biomarkers in the exosomes of their model cellular systems or patient biofluids. You will receive your data back in approximately 4-6 weeks. Nucleic Acid profiling and Next-Generation Sequencing Profiling of EV associated NA by qpcr. Next-Generation Sequencing (NGS) Profiling of RNA/DNA exosome markers. Probes can be designed by the customer and sent to HBM-LS. is performed using Ion Torrent and SOLiD System (Thermo Fisher Scientific) NGS platforms. 70
5 Catalog 2018 Section 7 Custom Product HansaBioMed Life Sciences can design and create a personalized kit by choosing from a wide variety of reagents and tools in our catalog. Product Custom ELISA immunoplate Custom immunobeads Custom product ELISA plates can be covalently coated with EV-binding antibodies. 1- Trasparent, white and black plates are available. 2- Coating antibody can be chosen by our antibody list (see section 6) or sent to HBM by customers. Latex immunobeads will be covalently coupled with the antibodies chosen from our antibody list, or sent to HBM-LS by customers. We provide the following product sizes: 1- Immunobeads for 10 reactions 2- Immunobeads for 20 reactions 3- Immunobeads for 50 reactions Customer can customize our ExoTEST platform chosing ELISA Custom ExoTEST plate and detection antibody (see section 3, Quantification kits) For other Custom made solutions and for information contact us at info@hansabiomed.eu or visit our website and project collaboration HansaBioMed Life Sciences is open for collaboration with private or academic partners in research activites related to different fields of exosome research and applications. Contact us for: and project collaboration We can provide biomolecule modifications such as: protein labeling with fluorescence dyes, conjugation of biomolecules with affinity ligands, PEGyation Biomolecule labeling and modification of biomolecules, immobilization of biomolecules on different surfaces (hydrogels, molded polymers, synthetic membranes) etc. Development of new assays for exosome marker identification Development of tools for capturing and enriching specific exosome subpopulations Projects and collaborations and technical consulting We can develop personalized ELISA assays for a specific exosome protein marker quantification or qpcr assays for identification of mirna and DNA markers. We can develop tools for capturing and isolating specific exosome subpopulations to improve comprehensive exosome profiling in diseases. Contact us for: OEM product development, cell free regenerative medicine, collaboration in research projects (Horizon 2020, Eurostars, NIH grants, etc). 71
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