Affiblot: a dot blot-based screening device for selection of reliable antibodies

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dc.contributor.author Svobodová, Zuzana
dc.contributor.author Novotný, Jakub
dc.contributor.author Ospálková, Barbora
dc.contributor.author Slováková, Marcela
dc.contributor.author Bílková, Zuzana
dc.contributor.author Foret, Frantisek
dc.date.accessioned 2022-06-03T12:11:56Z
dc.date.available 2022-06-03T12:11:56Z
dc.date.issued 2021
dc.identifier.issn 1759-9660
dc.identifier.uri https://hdl.handle.net/10195/79057
dc.description.abstract The key factor in the development of antibody-based assays is to find an antibody that has an appropriate affinity, high specificity, and low cross-reactivity. However, this task is not easy to carry out since the research antibodies on the market may suffer from low specificity and reproducibility. Here, we report on a palm-sized dot blot-based device, called the affiblot, that has a specially designed lid that allows simultaneous semi-quantitative comparison of up to five antibodies from different suppliers regarding their affinity/avidity, cross-reactivity, and batch-to-batch reliability. The only required peripheral equipment is a vacuum pump, a camera, and densitometry software. The affiblot device was tested for its functionality and its measurements were compared against those obtained by standard dot blot and ELISA. The benefit over these methods, when various antibodies are evaluated, is in its simplicity. It allows easy antigen deposition, fast application and the discarding of the solutions, a compact undivided membrane, and therefore significant decrease of labor. The device was tested with specific anti-ApoE, anti-EpCAM, anti-Salmonella, anti-E. coli, and anti-Listeria antibodies from different suppliers. Their properties were compared for their ability to interact specifically with antigen and/or non-target structures and the best-suited antibody for the intended application was identified. eng
dc.format p. 3874-3884 eng
dc.language.iso eng
dc.publisher Royal Society of Chemistry eng
dc.relation.ispartof Analytical Methods, volume 13, issue: 35 eng
dc.rights open access (green) eng
dc.subject avidity-Elisa eng
dc.subject validation eng
dc.subject specificity eng
dc.subject dot-blot eng
dc.subject validace cze
dc.subject dot-blot cze
dc.subject specifita cze
dc.subject avidita-ELISA cze
dc.title Affiblot: a dot blot-based screening device for selection of reliable antibodies eng
dc.title.alternative Affiblot: screeningové dot-blot zařízení pro selekci vhodných protilátek cze
dc.type article eng
dc.description.abstract-translated Klíčovým faktorem ve vývoji testů na bázi protilátek je najít protilátku, která má vhodnou afinitu, vysokou specificitu a nízkou zkříženou reaktivitu. To však není snadné provést, protože výzkumné protilátky na trhu mohou trpět nízkou specificitou a reprodukovatelností. Zde referujeme o zařízení na bázi dot blot velikosti dlaně, zvaném affiblot, které má speciálně navržené víko, které umožňuje simultánní semikvantitativní srovnání až pěti protilátek od různých dodavatelů, pokud jde o jejich afinitu/aviditu, zkříženou reaktivitu, a spolehlivost mezi jednotlivými dávkami. cze
dc.peerreviewed yes eng
dc.publicationstatus submitted version (preprint) eng
dc.identifier.doi 10.1039/d1ay00955a
dc.relation.publisherversion https://pubs.rsc.org/en/content/articlelanding/2021/AY/D1AY00955A
dc.project.ID EF17_048/0007421/NANOBIO - posilování mezioborové spolupráce ve výzkumu nanomateriálů a při studiu jejich účinků na živé organismy cze
dc.identifier.wos 000687267500001
dc.identifier.scopus 2-s2.0-85115395353
dc.identifier.obd 39886751


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