Research: PhDInstitut Curie · Translational Research DepartmentFrance · Paris
Article: 1(st) French-Israeli International Conference on B Cells and therapeutic antibodies: October 23-25, 2011 Jerusalem, Israel.Claude-Agnès Reynaud, Sandrine Moutel, Marie-Caroline Dieu-Nosjean, Reuven Laskov, Jean-Luc TeillaudmAbs 07/2012; 4(4):426-33.
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ABSTRACT: 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.Methods in molecular biology (Clifton, N.J.) 01/2012; 907:667-79.
Article: [Intrabodies, potent tools to unravel the function and dynamics of intracellular proteins].Sandrine Moutel, Franck Perez[show abstract] [hide abstract]
ABSTRACT: In the 1980s, progress in molecular biology enabled the manipulation and cloning of antibody fragments as functional scFv (single chain Fv). Because of their small size and relative ease of expression, scFv opened the road for new medical and biotechnological applications. scFvs can be easily expressed and targeted to different cellular compartments (cytosol, nucleus, endoplasmic reticulum, mitochondria, inner surface of the plasma membrane, etc.), using specific signals to target or retain them in a given compartment. Recombinant antibodies can thus be used as intracellular antibodies (intrabody) to neutralize, disrupt or track endogenous antigen. Intrabodies not only represent new tools for fundamental research to study the dynamics of endogenous proteins, but may also bring interesting options for applied research in terms of intracellular immunization for therapeutic use.Medecine sciences: M/S 12/2009; 25(12):1173-6. · 0.64 Impact Factor
Article: Direct selection of monoclonal phosphospecific antibodies without prior phosphoamino acid mapping.Ole Vielemeyer, Hebao Yuan, Sandrine Moutel, Rénette Saint-Fort, Danming Tang, Clément Nizak, Bruno Goud, Yanzhuang Wang, Franck Perez[show abstract] [hide abstract]
ABSTRACT: In the current post-genomic era, large scale efforts are underway to functionally explore the proteome by assembling large antibody libraries. However, because many proteins are modified post-translationally to regulate their function, collections of modification-specific sensors are also needed. Here we applied a novel approach to select monoclonal phosphospecific antibodies directly from the full-length protein and without up-front phosphoamino acid identification. We chose as antigen GRASP65, a well studied Golgi phosphoprotein. Bacterially produced full-length protein was first incubated with mitotic cytosol, thus allowing modification by naturally occurring kinases, and then used directly for affinity-based antibody selection using a single chain variable fragment phagemid library. In less than 1 week, three distinct and highly functional monoclonal phosphospecific antibodies against two GRASP65 epitopes were obtained and subsequently characterized. The presented approach is carried out fully in vitro, requires no prior knowledge of the phosphoamino acid identity, and is fast and inexpensive. It therefore has great potential to be an attractive alternative to classic animal-based protocols for the selection of post-translation modification sensors and thus to become an invaluable tool in our quest to understand the proteome in all its complexity.Journal of Biological Chemistry 07/2009; 284(31):20791-5. · 4.77 Impact Factor
Sandrine Moutel, Ahmed El Marjou, Ole Vielemeyer, Clément Nizak, Philippe Benaroch, Stefan Dübel, Franck Perez[show abstract] [hide abstract]
ABSTRACT: Genomic, transcriptomic and proteomic projects often suffer from a lack of functional validation creating a strong demand for specific and versatile antibodies. Antibody phage display represents an attractive approach to select rapidly in vitro the equivalent of monoclonal antibodies, like single chain Fv antibodies, in an inexpensive and animal free way. However, so far, recombinant antibodies have not managed to impose themselves as efficient alternatives to natural antibodies. We developed a series of vectors that allow one to easily fuse single chain Fv antibodies to Fc domains of immunoglobulins, improving their sensitivity and facilitating their use. This series enables the fusion of single chain Fv antibodies with human, mouse or rabbit Fc so that a given antibody is no longer restricted to a particular species. This opens up unlimited multiplexing possibilities and gives additional value to recombinant antibodies. We also show that this multi-Fc species production system can be applied to natural monoclonal antibodies cloned as single chain Fv antibodies and we converted the widely used 9E10 mouse anti-Myc-tag antibody into a human and a rabbit antibody. Altogether, this new expression system, that brings constant quality, sensitivity and unique versatility, will be important to broaden the use of recombinant and natural monoclonal antibodies both for laboratory and diagnosis use.BMC Biotechnology 03/2009; 9:14. · 2.35 Impact Factor