A cell ELISA based method for exosome detection in diagnostic and therapeutic applications
Objective Exosomes are nano-scaled carriers of miRNA, mRNA and proteins which are secreted from viable cells. Exosome detection within serum, saliva and semen offers diagnostic value for detection of various diseases including cancer. In the present study, we have lunched an in vitro study to develo...
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| Veröffentlicht in: | Biotechnology letters 2019-05, Vol.41 (4-5), p.523-531 |
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| creator | Khodashenas, Shabanali Khalili, Saeed Forouzandeh Moghadam, Mehdi |
| description | Objective
Exosomes are nano-scaled carriers of miRNA, mRNA and proteins which are secreted from viable cells. Exosome detection within serum, saliva and semen offers diagnostic value for detection of various diseases including cancer. In the present study, we have lunched an in vitro study to develop a more efficient method for exosome detection. In this regard, the recombinant LAMP-DARPins (capable of Her2 targeting) gene was packed within the lentiviral particles and stably transferred into the HEK293T cells. The morphology and sizes of the obtained exosomes were characterized by TEM and zeta sizer. The expression of LAMP-DARPins antigen on the exosome surface was verified by western blotting. Ultimately, the efficiency of cell surface ELISA in suspension method was examined for exosome detection.
Results
The exosome particles were successfully harvested and purified from transfected cells. The sizes of exosome particles were determined to be 90 nm using zeta sizer instrument. The TEM scan showed that the exosomes are cap like shaped and their sizes range between 40 and 150 nm. An observed 120 kDa band on western blotting paper indicated the LAMP-DARPins antigen expression on exosome surfaces. The results of cell surface ELISA in suspension method were superior to the results of conventional cell ELISA.
Conclusions
It could be concluded that the cell surface ELISA in suspension method could be an amenable method to detect exosome particles within the biological samples. Moreover, the method could be modified to evaluate the ability of exosomes to interact with target cells in both diagnostic and therapeutic applications. |
| doi_str_mv | 10.1007/s10529-019-02667-5 |
| format | Article |
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Exosomes are nano-scaled carriers of miRNA, mRNA and proteins which are secreted from viable cells. Exosome detection within serum, saliva and semen offers diagnostic value for detection of various diseases including cancer. In the present study, we have lunched an in vitro study to develop a more efficient method for exosome detection. In this regard, the recombinant LAMP-DARPins (capable of Her2 targeting) gene was packed within the lentiviral particles and stably transferred into the HEK293T cells. The morphology and sizes of the obtained exosomes were characterized by TEM and zeta sizer. The expression of LAMP-DARPins antigen on the exosome surface was verified by western blotting. Ultimately, the efficiency of cell surface ELISA in suspension method was examined for exosome detection.
Results
The exosome particles were successfully harvested and purified from transfected cells. The sizes of exosome particles were determined to be 90 nm using zeta sizer instrument. The TEM scan showed that the exosomes are cap like shaped and their sizes range between 40 and 150 nm. An observed 120 kDa band on western blotting paper indicated the LAMP-DARPins antigen expression on exosome surfaces. The results of cell surface ELISA in suspension method were superior to the results of conventional cell ELISA.
Conclusions
It could be concluded that the cell surface ELISA in suspension method could be an amenable method to detect exosome particles within the biological samples. Moreover, the method could be modified to evaluate the ability of exosomes to interact with target cells in both diagnostic and therapeutic applications.</description><identifier>ISSN: 0141-5492</identifier><identifier>EISSN: 1573-6776</identifier><identifier>DOI: 10.1007/s10529-019-02667-5</identifier><identifier>PMID: 30963341</identifier><language>eng</language><publisher>Dordrecht: Springer Netherlands</publisher><subject>Antigens ; Applied Microbiology ; Biochemistry ; Biological properties ; Biological samples ; Biomedical and Life Sciences ; Biotechnology ; Cancer ; Cell surface ; Diagnostic software ; Diagnostic systems ; Enzyme-linked immunosorbent assay ; ErbB-2 protein ; Exosomes ; Life Sciences ; Microbiology ; miRNA ; Morphology ; mRNA ; Original Research Paper ; Proteins ; Saliva ; Semen ; Therapeutic applications ; Western blotting</subject><ispartof>Biotechnology letters, 2019-05, Vol.41 (4-5), p.523-531</ispartof><rights>Springer Nature B.V. 2019</rights><rights>Biotechnology Letters is a copyright of Springer, (2019). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c478t-2dbbecc9c45be95ba5a994d78b50a81850c7125168f8443084595b9db2f8206b3</citedby><cites>FETCH-LOGICAL-c478t-2dbbecc9c45be95ba5a994d78b50a81850c7125168f8443084595b9db2f8206b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10529-019-02667-5$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10529-019-02667-5$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27903,27904,41467,42536,51297</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30963341$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Khodashenas, Shabanali</creatorcontrib><creatorcontrib>Khalili, Saeed</creatorcontrib><creatorcontrib>Forouzandeh Moghadam, Mehdi</creatorcontrib><title>A cell ELISA based method for exosome detection in diagnostic and therapeutic applications</title><title>Biotechnology letters</title><addtitle>Biotechnol Lett</addtitle><addtitle>Biotechnol Lett</addtitle><description>Objective
Exosomes are nano-scaled carriers of miRNA, mRNA and proteins which are secreted from viable cells. Exosome detection within serum, saliva and semen offers diagnostic value for detection of various diseases including cancer. In the present study, we have lunched an in vitro study to develop a more efficient method for exosome detection. In this regard, the recombinant LAMP-DARPins (capable of Her2 targeting) gene was packed within the lentiviral particles and stably transferred into the HEK293T cells. The morphology and sizes of the obtained exosomes were characterized by TEM and zeta sizer. The expression of LAMP-DARPins antigen on the exosome surface was verified by western blotting. Ultimately, the efficiency of cell surface ELISA in suspension method was examined for exosome detection.
Results
The exosome particles were successfully harvested and purified from transfected cells. The sizes of exosome particles were determined to be 90 nm using zeta sizer instrument. The TEM scan showed that the exosomes are cap like shaped and their sizes range between 40 and 150 nm. An observed 120 kDa band on western blotting paper indicated the LAMP-DARPins antigen expression on exosome surfaces. The results of cell surface ELISA in suspension method were superior to the results of conventional cell ELISA.
Conclusions
It could be concluded that the cell surface ELISA in suspension method could be an amenable method to detect exosome particles within the biological samples. Moreover, the method could be modified to evaluate the ability of exosomes to interact with target cells in both diagnostic and therapeutic applications.</description><subject>Antigens</subject><subject>Applied Microbiology</subject><subject>Biochemistry</subject><subject>Biological properties</subject><subject>Biological samples</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Cancer</subject><subject>Cell surface</subject><subject>Diagnostic software</subject><subject>Diagnostic systems</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>ErbB-2 protein</subject><subject>Exosomes</subject><subject>Life Sciences</subject><subject>Microbiology</subject><subject>miRNA</subject><subject>Morphology</subject><subject>mRNA</subject><subject>Original Research Paper</subject><subject>Proteins</subject><subject>Saliva</subject><subject>Semen</subject><subject>Therapeutic applications</subject><subject>Western blotting</subject><issn>0141-5492</issn><issn>1573-6776</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNp9kE1v1DAQhi0EotvCH-CALHHhEjp27Ng-rqoClVbiUHrpxfLHpE21iYOdSPDv8XZLkThwsEaWn3nH8xDyjsEnBqDOCwPJTQOsHt51qpEvyIZJ1TadUt1LsgEmWCOF4SfktJQHADAK1Gty0oLp2lawDbnd0oD7Pb3cXV1vqXcFIx1xuU-R9ilT_JlKGpFGXDAsQ5roMNE4uLsplWUI1E2RLveY3Yzr432e90NwB7K8Ia96ty_49qmekZvPl98vvja7b1-uLra7Jgill4ZH7zEEE4T0aKR30hkjotJegtNMSwiKcck63WshWtBCVspEz3vNofPtGfl4zJ1z-rFiWew4lMNSbsK0FssrxVvgTFf0wz_oQ1rzVH93oGTVU7VUih-pkFMpGXs752F0-ZdlYA_m7dG8rebto3kra9P7p-jVjxifW_6orkB7BEp9mu4w_539n9jfaUqNGQ</recordid><startdate>20190501</startdate><enddate>20190501</enddate><creator>Khodashenas, Shabanali</creator><creator>Khalili, Saeed</creator><creator>Forouzandeh Moghadam, Mehdi</creator><general>Springer Netherlands</general><general>Springer Nature 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cell ELISA based method for exosome detection in diagnostic and therapeutic applications</title><author>Khodashenas, Shabanali ; Khalili, Saeed ; Forouzandeh Moghadam, Mehdi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c478t-2dbbecc9c45be95ba5a994d78b50a81850c7125168f8443084595b9db2f8206b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Antigens</topic><topic>Applied Microbiology</topic><topic>Biochemistry</topic><topic>Biological properties</topic><topic>Biological samples</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Cancer</topic><topic>Cell surface</topic><topic>Diagnostic software</topic><topic>Diagnostic systems</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>ErbB-2 protein</topic><topic>Exosomes</topic><topic>Life Sciences</topic><topic>Microbiology</topic><topic>miRNA</topic><topic>Morphology</topic><topic>mRNA</topic><topic>Original Research Paper</topic><topic>Proteins</topic><topic>Saliva</topic><topic>Semen</topic><topic>Therapeutic applications</topic><topic>Western blotting</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Khodashenas, Shabanali</creatorcontrib><creatorcontrib>Khalili, Saeed</creatorcontrib><creatorcontrib>Forouzandeh Moghadam, Mehdi</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Chemoreception Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Health & Medical 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ELISA based method for exosome detection in diagnostic and therapeutic applications</atitle><jtitle>Biotechnology letters</jtitle><stitle>Biotechnol Lett</stitle><addtitle>Biotechnol Lett</addtitle><date>2019-05-01</date><risdate>2019</risdate><volume>41</volume><issue>4-5</issue><spage>523</spage><epage>531</epage><pages>523-531</pages><issn>0141-5492</issn><eissn>1573-6776</eissn><abstract>Objective
Exosomes are nano-scaled carriers of miRNA, mRNA and proteins which are secreted from viable cells. Exosome detection within serum, saliva and semen offers diagnostic value for detection of various diseases including cancer. In the present study, we have lunched an in vitro study to develop a more efficient method for exosome detection. In this regard, the recombinant LAMP-DARPins (capable of Her2 targeting) gene was packed within the lentiviral particles and stably transferred into the HEK293T cells. The morphology and sizes of the obtained exosomes were characterized by TEM and zeta sizer. The expression of LAMP-DARPins antigen on the exosome surface was verified by western blotting. Ultimately, the efficiency of cell surface ELISA in suspension method was examined for exosome detection.
Results
The exosome particles were successfully harvested and purified from transfected cells. The sizes of exosome particles were determined to be 90 nm using zeta sizer instrument. The TEM scan showed that the exosomes are cap like shaped and their sizes range between 40 and 150 nm. An observed 120 kDa band on western blotting paper indicated the LAMP-DARPins antigen expression on exosome surfaces. The results of cell surface ELISA in suspension method were superior to the results of conventional cell ELISA.
Conclusions
It could be concluded that the cell surface ELISA in suspension method could be an amenable method to detect exosome particles within the biological samples. Moreover, the method could be modified to evaluate the ability of exosomes to interact with target cells in both diagnostic and therapeutic applications.</abstract><cop>Dordrecht</cop><pub>Springer Netherlands</pub><pmid>30963341</pmid><doi>10.1007/s10529-019-02667-5</doi><tpages>9</tpages></addata></record> |
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| subjects | Antigens Applied Microbiology Biochemistry Biological properties Biological samples Biomedical and Life Sciences Biotechnology Cancer Cell surface Diagnostic software Diagnostic systems Enzyme-linked immunosorbent assay ErbB-2 protein Exosomes Life Sciences Microbiology miRNA Morphology mRNA Original Research Paper Proteins Saliva Semen Therapeutic applications Western blotting |
| title | A cell ELISA based method for exosome detection in diagnostic and therapeutic applications |
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