Anticorps thérapeutiques (TAB-IP) et anticorps recombinants

Publications de l’équipe

Année de publication : 2016

Sandrine Moutel, Nicolas Bery, Virginie Bernard, Laura Keller, Emilie Lemesre, Ario de Marco, Laetitia Ligat, Jean-Christophe Rain, Gilles Favre, Aurélien Olichon, Franck Perez (2016 Jul 20)

NaLi-H1: A universal synthetic library of humanized nanobodies providing highly functional antibodies and intrabodies.

eLife : DOI : 10.7554/eLife.16228 En savoir plus
Résumé

In vitro selection of antibodies allows to obtain highly functional binders, rapidly and at lower cost. Here, we describe the first fully synthetic phage display library of humanized llama single domain antibody (NaLi-H1: Nanobody Library Humanized 1). Based on a humanized synthetic single domain antibody (hs2dAb) scaffold optimized for intracellular stability, the highly diverse library provides high affinity binders without animal immunization. NaLi-H1 was screened following several selection schemes against various targets (Fluorescent proteins, actin, tubulin, p53, HP1). Conformation antibodies against active RHO GTPase were also obtained. Selected hs2dAb were used in various immunoassays and were often found to be functional intrabodies, enabling tracking or inhibition of endogenous targets. Functionalization of intrabodies allowed specific protein knockdown in living cells. Finally, direct selection against the surface of tumor cells produced hs2dAb directed against tumor-specific antigens further highlighting the potential use of this library for therapeutic applications.

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Année de publication : 2015

Marina Mikhaylova, Bas M C Cloin, Kieran Finan, Robert van den Berg, Jalmar Teeuw, Marta M Kijanka, Mikolaj Sokolowski, Eugene A Katrukha, Manuel Maidorn, Felipe Opazo, Sandrine Moutel, Marylin Vantard, Frank Perez, Paul M P van Bergen en Henegouwen, Casper C Hoogenraad, Helge Ewers, Lukas C Kapitein (2015 Aug 12)

Resolving bundled microtubules using anti-tubulin nanobodies.

Nature communications : 7933 : DOI : 10.1038/ncomms8933 En savoir plus
Résumé

Microtubules are hollow biopolymers of 25-nm diameter and are key constituents of the cytoskeleton. In neurons, microtubules are organized differently between axons and dendrites, but their precise organization in different compartments is not completely understood. Super-resolution microscopy techniques can detect specific structures at an increased resolution, but the narrow spacing between neuronal microtubules poses challenges because most existing labelling strategies increase the effective microtubule diameter by 20-40 nm and will thereby blend neighbouring microtubules into one structure. Here we develop single-chain antibody fragments (nanobodies) against tubulin to achieve super-resolution imaging of microtubules with a decreased apparent diameter. To test the resolving power of these novel probes, we generate microtubule bundles with a known spacing of 50-70 nm and successfully resolve individual microtubules. Individual bundled microtubules can also be resolved in different mammalian cells, including hippocampal neurons, allowing novel insights into fundamental mechanisms of microtubule organization in cell- and neurobiology.

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Année de publication : 2014

Selma Djender, Aurelie Schneider, Anne Beugnet, Ronan Crepin, Klervi Even Desrumeaux, Chiara Romani, Sandrine Moutel, Franck Perez, Ario de Marco (2014 Sep 17)

Bacterial cytoplasm as an effective cell compartment for producing functional VHH-based affinity reagents and Camelidae IgG-like recombinant antibodies.

Microbial cell factories : 140 : DOI : 10.1186/s12934-014-0140-1 En savoir plus
Résumé

The isolation of recombinant antibody fragments from displayed libraries represents a powerful alternative to the generation of IgGs using hybridoma technology. The selected antibody fragments can then be easily engineered into (multi)-tagged constructs of variable mass and complexity as well as reconstituted into Camelidae IgG-like molecules when expressed fused to Fc domains. Nevertheless, all antibody constructs depend on an oxidizing environment for correct folding and consequently still belong to the proteins difficult to express in bacteria. In such organisms they are mostly produced at low yields in the periplasmic space.

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Année de publication : 2012

Sandrine Moutel, Clément Nizak, Franck Perez (2012 Aug 22)

Selection and use of intracellular antibodies (intrabodies).

Methods in molecular biology (Clifton, N.J.) : 667-79 : DOI : 10.1007/978-1-61779-974-7_37 En savoir plus
Résumé

Intracellularly expressed recombinant antibodies, or intrabodies, are powerful tools for cell biology studies as well as therapeutic applications. Cell biologists use them to either block the intracellular antibody target or to image endogenous target dynamics. We describe here methods to select recombinant antibodies from antibody phage display libraries and to subsequently express them as fluorescent intrabodies.

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