Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy

Summary: Fast-dissociating, highly specific monoclonal antibodies (FDSAs) are single-molecule imaging probes useful for many biological assays including consecutive, multiplexable super-resolution microscopy. We developed a screening assay to characterize the kinetics of antibody-antigen interaction...

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Autores principales:	Takushi Miyoshi, Thomas B. Friedman, Naoki Watanabe
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Single-molecule Assays High Throughput Screening Microscopy Antibody Molecular/Chemical Probes Protein Biochemistry Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/e1fc8423d0574eea997bfe115e2c0405
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spelling	oai:doaj.org-article:e1fc8423d0574eea997bfe115e2c04052021-11-20T05:13:53ZFast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy2666-166710.1016/j.xpro.2021.100967https://doaj.org/article/e1fc8423d0574eea997bfe115e2c04052021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666166721006730https://doaj.org/toc/2666-1667Summary: Fast-dissociating, highly specific monoclonal antibodies (FDSAs) are single-molecule imaging probes useful for many biological assays including consecutive, multiplexable super-resolution microscopy. We developed a screening assay to characterize the kinetics of antibody-antigen interactions using single-molecule microscopy and established a pipeline to identify FDSAs from thousands of monoclonal candidates. Provided here are detailed protocols to prepare multi-well glass-bottom plates necessary for our assay to identify hybridoma clones secreting FDSAs. Synthesis of fluorescently labeled Fab fragments (Fab probes) from FDSAs is also described.For complete details on the use and execution of this protocol, please refer to Miyoshi et al. (2021).Takushi MiyoshiThomas B. FriedmanNaoki WatanabeElsevierarticleSingle-molecule AssaysHigh Throughput ScreeningMicroscopyAntibodyMolecular/Chemical ProbesProtein BiochemistryScience (General)Q1-390ENSTAR Protocols, Vol 2, Iss 4, Pp 100967- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Single-molecule Assays High Throughput Screening Microscopy Antibody Molecular/Chemical Probes Protein Biochemistry Science (General) Q1-390
spellingShingle	Single-molecule Assays High Throughput Screening Microscopy Antibody Molecular/Chemical Probes Protein Biochemistry Science (General) Q1-390 Takushi Miyoshi Thomas B. Friedman Naoki Watanabe Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy
description	Summary: Fast-dissociating, highly specific monoclonal antibodies (FDSAs) are single-molecule imaging probes useful for many biological assays including consecutive, multiplexable super-resolution microscopy. We developed a screening assay to characterize the kinetics of antibody-antigen interactions using single-molecule microscopy and established a pipeline to identify FDSAs from thousands of monoclonal candidates. Provided here are detailed protocols to prepare multi-well glass-bottom plates necessary for our assay to identify hybridoma clones secreting FDSAs. Synthesis of fluorescently labeled Fab fragments (Fab probes) from FDSAs is also described.For complete details on the use and execution of this protocol, please refer to Miyoshi et al. (2021).
format	article
author	Takushi Miyoshi Thomas B. Friedman Naoki Watanabe
author_facet	Takushi Miyoshi Thomas B. Friedman Naoki Watanabe
author_sort	Takushi Miyoshi
title	Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy
title_short	Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy
title_full	Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy
title_fullStr	Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy
title_full_unstemmed	Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy
title_sort	fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/e1fc8423d0574eea997bfe115e2c0405
work_keys_str_mv	AT takushimiyoshi fastdissociatingbuthighlyspecificantibodiesarenoveltoolsinbiologyespeciallyusefulformultiplexsuperresolutionmicroscopy AT thomasbfriedman fastdissociatingbuthighlyspecificantibodiesarenoveltoolsinbiologyespeciallyusefulformultiplexsuperresolutionmicroscopy AT naokiwatanabe fastdissociatingbuthighlyspecificantibodiesarenoveltoolsinbiologyespeciallyusefulformultiplexsuperresolutionmicroscopy
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Fast-dissociating but highly specific antibodies are novel tools in biology, especially useful for multiplex super-resolution microscopy

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