Institute of Chemistry & Biology of Membranes & Nano-objects • Bordeaux

LIA France-Japan GEM Réseau RMN Aquitain Master AC2QMAPS IPEP master UREKA UMT FOLIES INDIA
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Jean Duhamel, 4 april 2019, 11am, IECB Internal CBMN seminar 7 March 2019: « my thesis in 180 seconds » + new PhD students presentation Sandrine VIllette, 7 february 2019, 2pm, ENSCBP(B) Internal CBMN Seminar : newcomers in CBMN, 17th of January, 2pm Internal CBMN Seminar: A. Simon, 6 Dec. 2018 Marisela Velez, 28 november 2018, 2pm, IECB Marc Bramkamp, 16 november 2018, 2pm, IECB Kazushi Kinbara, 12 november 2018, 11am, IECB Internal CBMN Seminar: J. Lang, 8 November 2018 Internal CBMN Seminars: A. Baudin, 6 September 2018 Plateforme production de protéines, 5 juillet 2018, 14h, ENSTBB Patrick Trouillas, 21 June 2018, 2pm, ENSCBP Lucie Khemtemourian, 21 June 2018, 11am, IECB Jonathan Faherty, 7th June, 11 AM, IECB Yann Fichou, 3 May 2018, 2pm, IECB Internal CBMN seminars: A. Ciaccafava and G. Compain, 5 April 2018 Alain Roussel, 29/03/2018, 4 pm, ENSCBP Simon Poly, 22 march 2018, 2pm, ENSCBP J.Crassous, M. Sallé & A. Martinez, 18/12/2017, 10 am, IECB O.Reinaud & V. Artero, 7/12/2017, 10 am, IECB R. Dos santos Morais, 30/11/2017, 4 PM, ENSCBP Christian Griesinger, November 24th, 11am, IECB Takahiro Muraoka, 16/11/2017, 11am, ENSCBP Vladimir Torbeev, 9/11/2017, 2pm, ENSCBP M. D. Smith & T.Constantieux, 3/11/2017, 2h30 pm, IECB James Nowick, 15 september 2017, 11am, IECB Emmanuelle Thinon, 28 June 2017, 11 am, IECB Caroline Tokarski, 15 June 2017, 4pm, ENSCBP Christian Salesse, 8 June 2017, 4pm, ENSCBP Patrice Rey, 01 June 2017, 2h30 pm, ENSCBP Christina Sizun, 18 May 2017, 4 pm, ENSCBP Marisela Velez, 5 May 2017, 11am, IECB Iqbal Choudhary, 27 April 2017, 4 pm, ENSCBP Vincent Aucagne, 21 April 2017, 11 am, IECB Sophie Zinn-Justin, 14 April 2017, 11 am, IECB Cécile Feuillie, 16 February 2017, 4 PM, ENSCBP Félix M. Goñi, 8 december 2016, 4 pm, ENSCBP Félix M. Goñi, 17 november 2016, 4 pm, ENSCBP Carl Creutz, 20 october 2016, 4 pm, ENSCBP Diego Romero, 30 september 2016, 11 am, IECB A. Ciaccafava, 8 september 2016, 14h, ENSCBP A. Ramamoorthy, 16 June 2016, 16h, ENSCBP Alexandre de Brevern, 2 june 2016, ENSCBP Isabelle Landrieu, 26 May, 16h, ENSCBP Frances Sepavoric, 12 May, 16h, ENSCBP Thomas Pradeu, 7 April 2016, 9h, ENSCBP Françoise Argoul, 3 march 2016, 16h, ENSCBP Corinne Loutelier-Bourhis, 16/12/2015, 16h Aristotelis XENAKIS, 3 december 2015, 16h Fabian Kiessling, 26 november 2015, 11h Michaël Molinari, 20 november 2015, 11h Dipankar Das Sarma, 28 October 2015, 14h. Olivier Donard, 22 october 2015, 16h, ENSCBP E. Morvan & A.Grelard, 17/12/2015, 16h, ENSCBP Christophe Cullin, 10 september 2015, 14 h Brigitte Lindet, 18 June 2015, 16h, ENSCBP(B) Marion Decossas, 4 June 2015, 16h, ENSCBP(B) Pascale Schellenberger, 21 Mai 2015, 16h F. Leal-Calderon, 7 May 2015, 16h, ENSCBP(B) Ibrahim Abdulhalim, 9 April 2015, 14h, ENSCBP Manon Carré, 26 March 2015, 14h, ENSCBP(B) C. Bure & JM Schmitter, 19 March 2015, 16h T. Ogata & H. Ihara, 17 March 2015, 11h, IECB Banafshe Larijani, 12 March 2015, 16h, ENSCBP
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LIA France-Japan GEM Réseau RMN Aquitain Master AC2QMAPS IPEP master UREKA UMT FOLIES INDIA
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Jean Duhamel, 4 april 2019, 11am, I... Internal CBMN seminar 7 March 2019... Sandrine VIllette, 7 february 2019,... Internal CBMN Seminar : newcomers ... Internal CBMN Seminar: A. Simon, 6 ... Marisela Velez, 28 november 2018, 2... Marc Bramkamp, 16 november 2018, 2p... Kazushi Kinbara, 12 november 2018, ... Internal CBMN Seminar: J. Lang, 8 N... Internal CBMN Seminars: A. Baudin, ... Plateforme production de protéine... Patrick Trouillas, 21 June 2018, 2p... Lucie Khemtemourian, 21 June 2018, ... Jonathan Faherty, 7th June, 11 AM, ... Yann Fichou, 3 May 2018, 2pm, IECB Internal CBMN seminars: A. Ciaccafa... Alain Roussel, 29/03/2018, 4 pm, EN... Simon Poly, 22 march 2018, 2pm, ENS... J.Crassous, M. Sallé & A. Martinez... O.Reinaud & V. Artero, 7/12/2017, 1... R. Dos santos Morais, 30/11/2017, 4... Christian Griesinger, November 24th... Takahiro Muraoka, 16/11/2017, 11am,... Vladimir Torbeev, 9/11/2017, 2pm, E... M. D. Smith & T.Constantieux, 3/11/... James Nowick, 15 september 2017, 11... Emmanuelle Thinon, 28 June 2017, 11... Caroline Tokarski, 15 June 2017, 4p... Christian Salesse, 8 June 2017, 4pm... Patrice Rey, 01 June 2017, 2h30 pm,... Christina Sizun, 18 May 2017, 4 pm,... Marisela Velez, 5 May 2017, 11am, I... Iqbal Choudhary, 27 April 2017, 4 p... Vincent Aucagne, 21 April 2017, 11 ... Sophie Zinn-Justin, 14 April 2017, ... Cécile Feuillie, 16 February 2017,... Félix M. Goñi, 8 december 2016, 4... Félix M. Goñi, 17 november 2016, ... Carl Creutz, 20 october 2016, 4 pm,... Diego Romero, 30 september 2016, 11... A. Ciaccafava, 8 september 2016, 14... A. Ramamoorthy, 16 June 2016, 16h,... Alexandre de Brevern, 2 june 2016, ... Isabelle Landrieu, 26 May, 16h, ENS... Frances Sepavoric, 12 May, 16h, ENS... Thomas Pradeu, 7 April 2016, 9h, EN... Françoise Argoul, 3 march 2016, 1... Corinne Loutelier-Bourhis, 16/12/20... Aristotelis XENAKIS, 3 december 201... Fabian Kiessling, 26 november 2015,... Michaël Molinari, 20 november 2015... Dipankar Das Sarma, 28 October 2015... Olivier Donard, 22 october 2015, 16... E. Morvan & A.Grelard, 17/12/2015, ... Christophe Cullin, 10 september 201... Brigitte Lindet, 18 June 2015, 16h,... Marion Decossas, 4 June 2015, 16h, ... Pascale Schellenberger, 21 Mai 2015... F. Leal-Calderon, 7 May 2015, 16h, ... Ibrahim Abdulhalim, 9 April 2015, 1... Manon Carré, 26 March 2015, 14h, ... C. Bure & JM Schmitter, 19 March 20... T. Ogata & H. Ihara, 17 March 2015,... Banafshe Larijani, 12 March 2015, 1...
Events News
Patricia DULOR Anthony BOUTER Olivier LAMBERT Jochen LANG Pascale SUBRA-PATERNAULT Maria URDACI Marie-Christine DURRIEU Xavier SANTARELLI Gilles GUICHARD Yann FERRAND Isabelle BESTEL Juan ELEZGARAY Antoine LOQUET Sophie LECOMTE Reiko ODA Marc BONNEU

Romain Linares thesis defense, 2 december 2016, at 1h30 pm, ENSCBP, Building B



The defense will take place, the 2 december 2016, 1h30 pm, in the auditorium of building B of  the ENSCBP, allée Geoffroy Saint Hilaire, Pessac.

 

Phenotyping, quantification and isolation of extracellular vesicles from blood plasma using gold or magnetic nanoparticles conjugated to proteins.

 

Abstract

 

Extracellular vesicles (EVs) are submicrometric membrane vesicles found in body fluids and produced by cells in response to various stimuli. EVs are involved in numerous physiological processes but also in pathologies as cancers or cardiovascular diseases. Even if EVs are largely studied, our knowledge about them remains limited. This is mainly caused by the difficulties to characterize EVs and by the lack of standardized methods allowing their characterization. The first part of my PhD work focused on the development and optimization of a protein thiolation method. Antibodies modified to expose few thiols were conjugated to gold nanoparticles functionalized with maleimides. The binding of thiolated antibodies to gold nanoparticles was quantitatively studied and optimal conjugation conditions were determined using biochemical methods. The second part of my PhD work concerned the characterization of blood plasma EVs from healthy subjects using transmission electron microscopy (TEM). EVs morphology, size and phenotype were determined by cryo-TEM combined with labelling with protein-conjugated gold nanoparticles. The near-absolute quantification of blood plasma EVs was achieved using an original TEM method based on the direct sedimentation of EVs onto TEM grids. The third part of this study consisted in developing an EV isolation method using AnxA5-conjugated magnetic particles. Conditions allowing total extraction of blood plasma phosphatidylserine-exposing EVs were determined using flow cytometry (FC).

 

This study presents a detailed characterization of blood plasma EVs from healthy subjects and can serve as a reference for future studies on EVs contained in pathological plasmas or other body fluids.

 

 

(left), cryo-transmission electron microscopy image of two extracellular vesicles from blood plasma. The vesicle tagged "1" is strongly labeled with Annexin-A5 conjugated-gold nanoparticles. This indicates that this vesicle exposes phosphatidylserine (PS), a procoagulant phospholipid, contrary to the vesicle tagged "2". (right), Representation of an Annexin-A5 conjugated-gold nanoparticle, allowing the labeling of PS-exposing membranes in TEM.

 

Keywords : Extracellular vesicles, Blood plasma, Protein thiolation, Protein-conjugated gold nanoparticles, Immunogold labelling, Magnetic extraction, Annexin-A5, Phosphatidylserine, Electron microscopy, Flow cytometry

 

Supervision : Alain Brisson (a.brisson@cbmn.u-bordeaux.fr)

Team : Vésicules Extracellulaires et Réparation Membranaire (A.Brisson)

 

 

 

 

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