Selection of functional single domain antibody fragments for interfering with protein-protein interactions inside cells: a "one plasmid" mammalian two-hybrid system.

As a complement to the intracellular antibody capture method to isolate intracellular single domain antibody fragments (iDabs) from high diverse libraries, we describe here a simple mammalian two-hybrid (M2H) protocol using a "bait-prey hybrid single plasmid" to assess those interfering iD...

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Номзүйн дэлгэрэнгүй
Үндсэн зохиолчид: Tanaka, T, Rabbitts, T
Формат: Book section
Хэвлэсэн: 2012
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author Tanaka, T
Rabbitts, T
author_facet Tanaka, T
Rabbitts, T
author_sort Tanaka, T
collection OXFORD
description As a complement to the intracellular antibody capture method to isolate intracellular single domain antibody fragments (iDabs) from high diverse libraries, we describe here a simple mammalian two-hybrid (M2H) protocol using a "bait-prey hybrid single plasmid" to assess those interfering iDabs that will block protein-protein interactions of a target with its natural partner proteins. This rapid method identifies interfering iDabs in one step and improves the reproducibility of the results between experiments and samples (e.g., different single domain antibody clones) compared to traditional M2H. This method yields functional, interfering iDabs and can be applied to any interfering molecule for use as a research tool or template for clinical inhibitor production.
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institution University of Oxford
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publishDate 2012
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spelling oxford-uuid:f5ea8e1f-4b1f-4030-b28c-1fbf7a3fbf1d2022-03-27T12:30:59ZSelection of functional single domain antibody fragments for interfering with protein-protein interactions inside cells: a "one plasmid" mammalian two-hybrid system.Book sectionhttp://purl.org/coar/resource_type/c_3248uuid:f5ea8e1f-4b1f-4030-b28c-1fbf7a3fbf1dSymplectic Elements at Oxford2012Tanaka, TRabbitts, TAs a complement to the intracellular antibody capture method to isolate intracellular single domain antibody fragments (iDabs) from high diverse libraries, we describe here a simple mammalian two-hybrid (M2H) protocol using a "bait-prey hybrid single plasmid" to assess those interfering iDabs that will block protein-protein interactions of a target with its natural partner proteins. This rapid method identifies interfering iDabs in one step and improves the reproducibility of the results between experiments and samples (e.g., different single domain antibody clones) compared to traditional M2H. This method yields functional, interfering iDabs and can be applied to any interfering molecule for use as a research tool or template for clinical inhibitor production.
spellingShingle Tanaka, T
Rabbitts, T
Selection of functional single domain antibody fragments for interfering with protein-protein interactions inside cells: a "one plasmid" mammalian two-hybrid system.
title Selection of functional single domain antibody fragments for interfering with protein-protein interactions inside cells: a "one plasmid" mammalian two-hybrid system.
title_full Selection of functional single domain antibody fragments for interfering with protein-protein interactions inside cells: a "one plasmid" mammalian two-hybrid system.
title_fullStr Selection of functional single domain antibody fragments for interfering with protein-protein interactions inside cells: a "one plasmid" mammalian two-hybrid system.
title_full_unstemmed Selection of functional single domain antibody fragments for interfering with protein-protein interactions inside cells: a "one plasmid" mammalian two-hybrid system.
title_short Selection of functional single domain antibody fragments for interfering with protein-protein interactions inside cells: a "one plasmid" mammalian two-hybrid system.
title_sort selection of functional single domain antibody fragments for interfering with protein protein interactions inside cells a one plasmid mammalian two hybrid system
work_keys_str_mv AT tanakat selectionoffunctionalsingledomainantibodyfragmentsforinterferingwithproteinproteininteractionsinsidecellsaoneplasmidmammaliantwohybridsystem
AT rabbittst selectionoffunctionalsingledomainantibodyfragmentsforinterferingwithproteinproteininteractionsinsidecellsaoneplasmidmammaliantwohybridsystem