Nanobody

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Nanobody

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Author : Ulrich Rothbauer
language : en
Publisher: MDPI
Release Date : 2021-03-30

Nanobody written by Ulrich Rothbauer and has been published by MDPI this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021-03-30 with Science categories.

Nanobodies have become outstanding tools for biomedical research, diagnostics and therapy. Recent advances in the identification and functionalization of target-specific nanobodies now make nanobody-based approaches broadly available to many researches in the field. This book provides a compilation of original research articles and comprehensive reviews covering important and up to date aspects of research on nanobodies and their applications for immunoassays, proteomics, protein crystallization and in vitro and in vivo imaging.

Rapid Nanobody Discovery And Novel Nanobody Engineering Strategies For The Study Of The Nuclear Pore Complex

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Author : Tino Pleiner
language : en
Publisher: Cuvillier Verlag
Release Date : 2017-04-03

Rapid Nanobody Discovery And Novel Nanobody Engineering Strategies For The Study Of The Nuclear Pore Complex written by Tino Pleiner and has been published by Cuvillier Verlag this book supported file pdf, txt, epub, kindle and other format this book has been release on 2017-04-03 with Science categories.

Nanobodies are recombinant single-domain antibodies of camelid origin. We set out to generate nanobodies as tools to study nuclear pore complexes (NPCs) that constitute the major passageways for nucleocytoplasmic exchange. Therefore, we developed an optimized phage display-based workflow for the generation of nanobodies from an immunized animal. The identified nanobodies were functionally expressed in the E. coli cytoplasm in a disulfide bond-free form, thus overcoming the engineering limitations imposed by conventional periplasmic secretion. Using protease-cleavable affinity tags, we developed a generic strategy for native protein complex isolation from eukaryotic sources, e.g. for structural analyses by electron microscopy. We further established a reliable fluorescent labeling strategy of nanobodies that is based on engineered surface cysteines and maleimide chemistry. In contrast to NHS labeling of lysines, maleimide labeling is site-specific, does not interfere with antigen recognition and yields superior imaging reagents, especially for super-resolution microscopy. Using our optimized workflow, we obtained a large toolbox of nanobodies against constituents of Xenopus NPCs that can now be used to study various unresolved aspects of NPC structure, assembly and function.

Nanobody

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Author : Ulrich Rothbauer
language : en
Publisher:
Release Date : 2021

Nanobody written by Ulrich Rothbauer and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021 with categories.

Nanobody Based Cancer Immunotherapy And Immunoimaging

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Author : Zahra Sharifzadeh
language : en
Publisher: Frontiers Media SA
Release Date : 2023-06-07

Nanobody Based Cancer Immunotherapy And Immunoimaging written by Zahra Sharifzadeh and has been published by Frontiers Media SA this book supported file pdf, txt, epub, kindle and other format this book has been release on 2023-06-07 with Science categories.

Engineering Nanobody Intracellular Behavior Through Framework Mutagenesis

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Author : John Grether Dingus
language : en
Publisher:
Release Date : 2021

Engineering Nanobody Intracellular Behavior Through Framework Mutagenesis written by John Grether Dingus and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021 with Immunoglobulins categories.

Antibodies and their derived fragments have traditionally been used to probe, neutralize, or interrogate primarily extracellular targets in living systems. While efforts have been taken to adapt the antigen-binding portions of conventional antibodies to the intracellular environment, a lack of consistent stable and soluble behavior has hampered their utility for intracellular studies in live cells. Nanobodies, variable domains derived from a special class of antibody lacking light chains, hold promise as intracellularly functional tools. At less than one-tenth the size of a conventional antibody, nanobodies retain full antigen binding function, and exhibit superior expressibility in the reducing environment of the cytoplasm. Furthermore, previous work in the Cepko lab has illustrated that nanobodies can be engineered to rely on antigen interaction for stability, a property facilitating the design of conditionally active tools that only function in cells expressing specific targets. In this dissertation, the generality of both intracellular stability and engineered, conditional instability are examined. In chapter 2, I highlight intracellular stability differences observed across a broad repertoire of previously identified nanobodies, interrogate sequence differences between intracellularly stable and unstable nanobodies, and develop a general approach to mutationally stabilize previously unstable nanobodies for intracellular expression. In chapter 3, I propose specific framework mutations, identified by mutational screens over nanobodies exhibiting disparate modes of target interaction, that facilitate both destabilization in the absence of antigen, and re-stabilization in the presence of antigen, transmissible across a broad range of nanobodies. This work will streamline the design and function of target-specific, intracellularly active protein tools.

Developing Antibody And Nanobody Based Platforms Technologies For Targeting The Tumor Microenvironment

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Author : Joseph P. Gallant
language : en
Publisher:
Release Date : 2023

Developing Antibody And Nanobody Based Platforms Technologies For Targeting The Tumor Microenvironment written by Joseph P. Gallant and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2023 with categories.

The development of both traditional antibodies and nanobodies in parallel allows for constructs with tunable pharmacokinetics to be employed in various applications. Here a full panel of antibodies and nanobody based constructs have are described for targeting an important tumor microenvironment antigen, fibroblast activation protein alpha. The traditional antibody described here has been evaluated as a potential PET/CT imaging agent and platform for an antibody drug conjugate for preclinical models of metastatic castration resistant prostate cancer. While the panel of nanobody based constructs ranging from a small monovalent camelid nanobody to a camelid nanobody Fc fusion were evaluate as PET/CT imaging agents and compared to relevant small molecule inhibitor-based tracers. The full range of biologics described and characterized in this work ranges from 12kDa to 155kDa and exemplifies the dynamic pharmacokinetics of biologics of varying sizes. These constructs may have broad applications as potent theranostic platforms.

A Nanobody Suite For Yeast Scaffold Nucleoporins Provides Details Of The Y Complex Structure And Nuclear Pore Complex Assembly

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Author : Sarah Ann Nordeen
language : en
Publisher:
Release Date : 2020

A Nanobody Suite For Yeast Scaffold Nucleoporins Provides Details Of The Y Complex Structure And Nuclear Pore Complex Assembly written by Sarah Ann Nordeen and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2020 with categories.

Using Data To Characterise The Relationship Between Nanobody Sequence Structure And Function

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Author : Laura Sophie Mitchell
language : en
Publisher:
Release Date : 2019

Using Data To Characterise The Relationship Between Nanobody Sequence Structure And Function written by Laura Sophie Mitchell and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019 with categories.

A Toolkit To Study Nuclear Transport Pathways Nanobody Mediated Inhibition Of Importin Related Nuclear Transport Receptors Ntrs

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Author : Dilantha Perera
language : en
Publisher: Cuvillier Verlag
Release Date : 2023-08-07

A Toolkit To Study Nuclear Transport Pathways Nanobody Mediated Inhibition Of Importin Related Nuclear Transport Receptors Ntrs written by Dilantha Perera and has been published by Cuvillier Verlag this book supported file pdf, txt, epub, kindle and other format this book has been release on 2023-08-07 with Science categories.

Eukaryotic cells are divided into a nuclear and a cytoplasmic compartment. This separates transcription from translation and makes gene expression dependent on nucleocytoplasmic transport. The members of the importin β superfamily function as shuttling nuclear transport receptors (NTRs) that recognize and actively transport cargoes through nuclear pores. An estimated 5 000 to 10 000 different human proteins are subject to active nuclear transport. Numerous cargo/NTR pairs have been identified, however, we are still far from a complete understanding as it has been very challenging to setup a systematic in vivo analysis that integrates the impact of all transport pathways. In this study, we obtained anti-NTR nanobodies against TRN1, Xpo4, Xpo7, and CAS. Our aim was to identify nanobodies, and prepare nanobody fusions, that impede nuclear pore-passage of the targeted NTR and thus, interrupt a given transport cycle. These nanobody fusions were observed to inhibit the partition of NTR/cargo complexes into a reconstituted FG phase. We also observed that the nanobodies and nanobody fusions inhibit NTR transport in permeabilized cells.

Nanobody Based Approaches For The Therapeutic Targeting Of Conserved T Cell Subsets In Cancer

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Author :
language : en
Publisher:
Release Date : 2017

Nanobody Based Approaches For The Therapeutic Targeting Of Conserved T Cell Subsets In Cancer written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2017 with categories.

Nanobody

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