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© Matteo Bonazzi, Edith Gouin
Observation en immunofluorescence d'une cellule infectée par Listeria monocytogenes. En bleu: marquage des protéines de surface de Listeria qui permet de visualiser les bactéries. En rouge et vert: marquage de l'actine, une protéine qui forme le cytosquelette des cellules. Les Listeria utilisent l'actine cellulaire pour former des "comêtes" et se déplacer à l'intérieur des cellules qu'elles infectent. Cell infected by Listeria monocytogenes. The surface proteins (in blue) of Listeria enable us to view the bacteria. Actin, a constituent protein of cells, is shown in red and green.
Project

DROPmAbs – Deep-mining of antibody repertoires using droplet-based microfluidics combined with barcoded deep sequencing for diagnostics and therapeutic antibody discovery

Starting Date
01
Nov 2014
Ending Date
31
Oct 2018
Status
Ongoing
Members
5
Structures
1

About

 In this project we propose to combine ultrahigh-throughput screening of primary B cells using droplet based microfluidics with next-generation sequencing to profile entire B cell antibody repertoires. This process will enable highly-efficient and rapid characterisation of antigen-specific B cells and establish an “anatomical map” of their distribution. We will couple droplet-based microfluidic screening based on antigen binding to a novel next-generation sequencing approach based on microfluidic DNA barcoding to retrieve the antibody sequences from each B cell specifically, whilst maintaining the information about the pairing of the heavy chain and light chain variable regions (VH and VL) and associating it with the affinity of each antibody for the antigen. We believe the combination of these two techniques will constitute a powerful tool for the screening of non-immortalized B cells and allow unprecedented deep-mining of antibody repertoires (since millions of B cells can be screened and sequenced per day) with multiple applications, including (i) fundamental research into idiopathic disease, (ii) diagnosis/prognosis of idiopathic disease (iii) vaccination and (iv) discovery of therapeutic antibodies. Applying this technology, we should be able to characterize the antibody repertoire in detail (the ‘antibodyome’) and generate an anatomical map of antibody response with respect to antigen nature, immunization route and B cell clone dissemination. This will be achieved via immunizations of mice by various routes as well as with a preclinical autoimmune model. Finally, we propose to demonstrate the pathogenic nature of antibodies in a given human pathological condition (allergic shock) as a proof of concept for establishing diagnostic screens in idiopathic diseases to which specific antibodies could be triggers or effectors.

Consortium:                                                                                                                                                                                                                           Antibodies in Therapy & Pathology, Institut Pasteur; Dr Pierre Bruhns (coordinator)                                                                       Biochemistry Laboratory, ESPCI-ParisTech; Pr Andrew Griffiths                                                                                                                            HiFiBiO SAS, ESPCI-ParisTech; Dr Allan Jensen                                                                                                                                                           Antibody engineering platform (PFIA), Institut Pasteur; Dr Pierre Lafaye

Fundings