Purification-independent immunoreagents obtained by displaying nanobodies on bacteria surface
The availability of preimmune libraries of antibody fragments allows for the fast generation of binders which can be expressed in both eukaryotic and prokaryotic systems. We exploited the recombinant nature of antibody fragments to demonstrate the possibility of expressing them as functional protein...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2019-06, Vol.103 (11), p.4443-4453 |
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| creator | Oloketuyi, Sandra Dilkaute, Carina Mazzega, Elisa Jose, Joachim de Marco, Ario |
| description | The availability of preimmune libraries of antibody fragments allows for the fast generation of binders which can be expressed in both eukaryotic and prokaryotic systems. We exploited the recombinant nature of antibody fragments to demonstrate the possibility of expressing them as functional proteins displayed on the surface of
Escherichia coli
and by such a way to generate living reagents ready-to-use for diagnostics. Such immunoreagents were effectively exploited without the necessity of any purification step to prepare immunocapture surfaces suitable for the diagnostic of both cancer cells and toxic microalgae. The same nanobody-displaying bacteria were also engineered to coexpress GFP in their cytoplasm. Suspensions of such living fluorescent immunoreagents effectively bound to eukaryotic cells making them visible and quantifiable by flow cytometry analysis and using 96-well plate readers. The collected data showed the suitability of such living immunoreagents for reproducible and inexpensive diagnostic applications. |
| doi_str_mv | 10.1007/s00253-019-09823-x |
| format | Article |
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Escherichia coli
and by such a way to generate living reagents ready-to-use for diagnostics. Such immunoreagents were effectively exploited without the necessity of any purification step to prepare immunocapture surfaces suitable for the diagnostic of both cancer cells and toxic microalgae. The same nanobody-displaying bacteria were also engineered to coexpress GFP in their cytoplasm. Suspensions of such living fluorescent immunoreagents effectively bound to eukaryotic cells making them visible and quantifiable by flow cytometry analysis and using 96-well plate readers. The collected data showed the suitability of such living immunoreagents for reproducible and inexpensive diagnostic applications.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-019-09823-x</identifier><identifier>PMID: 30989251</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Algae ; Antibiotics ; Antibodies ; Aquatic microorganisms ; Bacteria ; Binders ; Biomedical and Life Sciences ; Biotechnologically Relevant Enzymes and Proteins ; Biotechnology ; Cancer ; Cytoplasm ; Dehydrogenases ; Diagnostic software ; Diagnostic systems ; E coli ; Escherichia coli ; Flow cytometry ; Fluorescence ; Fragments ; Life Sciences ; Microbial Genetics and Genomics ; Microbiology ; Nanobodies ; Nanotechnology ; Observations ; Plasmids ; Proteins ; Purification ; Reagents ; Technology application</subject><ispartof>Applied microbiology and biotechnology, 2019-06, Vol.103 (11), p.4443-4453</ispartof><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2019</rights><rights>COPYRIGHT 2019 Springer</rights><rights>Applied Microbiology and Biotechnology is a copyright of Springer, (2019). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c513t-f387344f6109d2266bb14ca83794ab05ffbb479d39a1a0705babdcc194e4921b3</citedby><cites>FETCH-LOGICAL-c513t-f387344f6109d2266bb14ca83794ab05ffbb479d39a1a0705babdcc194e4921b3</cites><orcidid>0000-0001-7729-819X</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-019-09823-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-019-09823-x$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30989251$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Oloketuyi, Sandra</creatorcontrib><creatorcontrib>Dilkaute, Carina</creatorcontrib><creatorcontrib>Mazzega, Elisa</creatorcontrib><creatorcontrib>Jose, Joachim</creatorcontrib><creatorcontrib>de Marco, Ario</creatorcontrib><title>Purification-independent immunoreagents obtained by displaying nanobodies on bacteria surface</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>The availability of preimmune libraries of antibody fragments allows for the fast generation of binders which can be expressed in both eukaryotic and prokaryotic systems. We exploited the recombinant nature of antibody fragments to demonstrate the possibility of expressing them as functional proteins displayed on the surface of
Escherichia coli
and by such a way to generate living reagents ready-to-use for diagnostics. Such immunoreagents were effectively exploited without the necessity of any purification step to prepare immunocapture surfaces suitable for the diagnostic of both cancer cells and toxic microalgae. The same nanobody-displaying bacteria were also engineered to coexpress GFP in their cytoplasm. Suspensions of such living fluorescent immunoreagents effectively bound to eukaryotic cells making them visible and quantifiable by flow cytometry analysis and using 96-well plate readers. The collected data showed the suitability of such living immunoreagents for reproducible and inexpensive diagnostic applications.</description><subject>Algae</subject><subject>Antibiotics</subject><subject>Antibodies</subject><subject>Aquatic microorganisms</subject><subject>Bacteria</subject><subject>Binders</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnologically Relevant Enzymes and Proteins</subject><subject>Biotechnology</subject><subject>Cancer</subject><subject>Cytoplasm</subject><subject>Dehydrogenases</subject><subject>Diagnostic software</subject><subject>Diagnostic systems</subject><subject>E coli</subject><subject>Escherichia coli</subject><subject>Flow cytometry</subject><subject>Fluorescence</subject><subject>Fragments</subject><subject>Life Sciences</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Nanobodies</subject><subject>Nanotechnology</subject><subject>Observations</subject><subject>Plasmids</subject><subject>Proteins</subject><subject>Purification</subject><subject>Reagents</subject><subject>Technology 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biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Oloketuyi, Sandra</au><au>Dilkaute, Carina</au><au>Mazzega, Elisa</au><au>Jose, Joachim</au><au>de Marco, Ario</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Purification-independent immunoreagents obtained by displaying nanobodies on bacteria surface</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2019-06-01</date><risdate>2019</risdate><volume>103</volume><issue>11</issue><spage>4443</spage><epage>4453</epage><pages>4443-4453</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>The availability of preimmune libraries of antibody fragments allows for the fast generation of binders which can be expressed in both eukaryotic and prokaryotic systems. We exploited the recombinant nature of antibody fragments to demonstrate the possibility of expressing them as functional proteins displayed on the surface of
Escherichia coli
and by such a way to generate living reagents ready-to-use for diagnostics. Such immunoreagents were effectively exploited without the necessity of any purification step to prepare immunocapture surfaces suitable for the diagnostic of both cancer cells and toxic microalgae. The same nanobody-displaying bacteria were also engineered to coexpress GFP in their cytoplasm. Suspensions of such living fluorescent immunoreagents effectively bound to eukaryotic cells making them visible and quantifiable by flow cytometry analysis and using 96-well plate readers. The collected data showed the suitability of such living immunoreagents for reproducible and inexpensive diagnostic applications.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>30989251</pmid><doi>10.1007/s00253-019-09823-x</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0001-7729-819X</orcidid></addata></record> |
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| ispartof | Applied microbiology and biotechnology, 2019-06, Vol.103 (11), p.4443-4453 |
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| source | SpringerLink Journals - AutoHoldings |
| subjects | Algae Antibiotics Antibodies Aquatic microorganisms Bacteria Binders Biomedical and Life Sciences Biotechnologically Relevant Enzymes and Proteins Biotechnology Cancer Cytoplasm Dehydrogenases Diagnostic software Diagnostic systems E coli Escherichia coli Flow cytometry Fluorescence Fragments Life Sciences Microbial Genetics and Genomics Microbiology Nanobodies Nanotechnology Observations Plasmids Proteins Purification Reagents Technology application |
| title | Purification-independent immunoreagents obtained by displaying nanobodies on bacteria surface |
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