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Development of Host Cell Protein Impurities Quantification Methods by Mass Spectrometry to Control the Quality of Biopharmaceuticals Gauthier Husson

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Development of Host Cell Protein Impurities Quantification Methods by Mass Spectrometry to Control the Quality of Biopharmaceuticals Gauthier Husson Development of host cell protein impurities quantification methods by mass spectrometry to control the quality of biopharmaceuticals Gauthier Husson To cite this version: Gauthier Husson. Development of host cell protein impurities quantification methods by mass spec- trometry to control the quality of biopharmaceuticals. Analytical chemistry. Université de Strasbourg, 2017. English. NNT : 2017STRAF066. tel-01730667 HAL Id: tel-01730667 https://tel.archives-ouvertes.fr/tel-01730667 Submitted on 13 Mar 2018 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. UNIVERSITÉ DE STRASBOURG ÉCOLE DOCTORALE DES SCIENCES CHIMIQUES UMR 7178 THÈSE présentée par : Gauthier HUSSON soutenue le : 10 novembre 2017 pour obtenir le grade de : Docteur de l’université de Strasbourg Discipline / Spécialité : Chimie / Chimie Analytique DEVELOPMENT OF HOST CELL PROTEIN IMPURITIES QUANTIFICATION METHODS BY MASS SPECTROMETRY TO CONTROL THE QUALITY OF BIOPHARMACEUTICALS THÈSE dirigée par : Dr. VAN DORSSELAER Alain Directeur de recherche, CNRS, Université de Strasbourg, France Pr. BRACEWELL Daniel Professor, University College London, United Kingdom RAPPORTEURS : Dr. FERRO Myriam Directeur de recherche, CEA, Grenoble, France Dr. O’HARA John Director, Characterisation, UCB, Slough, United Kingdom AUTRES MEMBRES DU JURY : Dr. GERVAIS Annick Director PCMD I, ASB, UCB, Braine l’Alleud, Belgium Dr. CARAPITO Christine Chargée de recherche, CNRS, Université de Strasbourg, France A mes parents, Mon frère, Ma famille, Mes amis, « Le doute est le plus sûr instrument pour la découverte de la vérité ; c'est un verre brut qu'on donne à polir au temps et au génie pour aider tous les yeux à voir enfin les objets tels qu'ils sont » Joseph Michel Antoine Servan Acknowledgments Tout d’abord, je tiens à remercier Alain Van Dorsselaer et Sarah Cianférani pour m’avoir accueilli au Laboratoire de Spectrométrie de Masse Bio-Organique. Je voudrais remercier mes encadrants: un énorme merci Christine Carapito, pour tout ce que tu m’as appris, pour ton soutien au quotidien, pour ces discussions toujours enrichissantes. Merci pour ton implication, pour ta gentillesse. Merci de m’avoir fait découvrir et aimer la spectrométrie de masse. Merci Alain Van Dorsselaer pour vos précieux conseils tout au long de ma thèse. Un tout grand merci à Aurélie Delangle et Annick Gervais pour votre confiance, votre encadrement, vos réponses à mes nombreuses questions, votre soutien. Merci de me faire encore confiance aujourd’hui. A big thank you to Dan Bracewell, for your supervision at University College London, and your precious advices. I learned a lot during these five months, including mAb production and purification, but also English! Thank you to John O’Hara and Myriam Ferro for accepting to review and evaluate my work. Je voudrais remercier ceux avec qui j’ai collaboré durant ma thèse pour différents projets: Bruno Maucourt, Sabrina Bibi-Triki, Françoise Bringel, Muriel Bonnet et Sylvain Debard. Thank you to my colleagues from UCL: Hai Yuan, thanks for all the time we spent together in the lab, for the discussions, the brainstorming, your kindness. Sushobhan, or Shubho, I was so lucky to have you in the lab. You were my personal teacher, and I really enjoyed learning so many things from you. Damiano, thanks for having welcoming me in the team, learning me suspension cell culture and for all the after work sessions. Alex, mon soutien francophone parmi tous ces anglophones, merci pour ta bonne humeur, et ton soutien tout au long de mon séjour à l’UCL. Thanks to my colleagues from UCB: Matthew Hinchliffe, thank you for your strong support and all the advices regarding the cell culture and mAb purification. Thank you John O’Hara for your support, and your implication in the project. Merci Anne-Sophie pour ton aide concernant la recherche bibliographique. Merci à mes collègues du LSMBO, où j’ai passé la majeure partie de ma thèse, pour votre gentillesse et votre bonne humeur. Merci aux anciens: Diego, Gilles, Marine, merci pour votre accueil et vos conseils. Johann et Guillaume, merci pour vos réponses à mes nombreuses questions, et pour votre humour fort belge ! Merci Benoît pour ton humour un peu fou (et un peu belge aussi), et pour m’avoir accompagné au RU jusqu’au bout. Georg, merci pour ta gentillesse, ton humour, et tout ce que tu m’as appris lors de nos discussions scientifiques tardives ! Angela te passe le bonjour depuis l’espace ;) Merci Maurane pour avoir partagé une partie de ma thèse (je me souviendrai longtemps de ce développement de méthode SRM !). Merci Hélène, Agnès, Véronique, Fabrice Varrier, Fabrice Bertile, Luc, Alfred, Laurence, Christine Schaffer, Martine et Stella pour votre bonne humeur. Merci Danièle pour ta gentillesse cachée derrière cette façade alsacienne ! François, merci pour ta bonne humeur et pour tes blagues, j’espère que tu as commandé un chargeur de téléphone pour Noël Merci aux informaticiens pour leur aide au quotidien, Alex (vraiment merci pour ton aide précieuse à chaque fois que j’en avais besoin), Alex 2, Aymen, Patrick. Jean-Marc, merci pour tout ce que tu m’as appris, pour ton aide tout au long de ma thèse, pour ta bienveillance envers tous les p’tits jeunes ! Merci aussi pour ta bonne humeur. Le labo a de la chance de t’avoir ! Prends soin de Nelson ;) Merci aux supramol : Anthony et Thomas, bon courage pour vos thèses propres maggle ! Oscar, j’espère qu’un jour tu trouveras un sac à dos aussi beau que le mien. Maxime B, j’ai vraiment aimé discuter avec toi de science comme d’autres choses (« mets-toi donc à ma place, Stephen ! »), ces macdo tardifs, ces parties de pétanque, … Bon courage pour la fin de ta thèse, tes problèmes de chaleurs et d’humidité (je parle bien sûr du tiroir du robot). Stéphane, prends soin de ma co-équipière, et Steeve, bonne continuation dans ton nouveau bureau ! Merci Ludivine, Paola, Justine, Pauline, Blandine, Leslie et Nicolas pour votre gentillesse et votre bonne humeur. Paola et Blandine, bon courage pour vos thèses ! Justine, bon courage pour le Triple TOF Leslie, je me souviens de ce retour de SFEAP plutôt rigolo ! Félicitations pour ta thèse que tu viens de soutenir. Nicolas, bon courage pour ta thèse (écoute bien tout ce que te dit Jojo !). Merci Maxime E, « mon père », pour ta bonne humeur, et ton aide sur le traitement de données DIA… Ce n’était pas évident mais tu t’en es bien sorti, et je te souhaite le meilleur pour ta thèse à Maastricht. Merci Sebastian, pour tout ce que tu m’as appris. Je ne connaissais rien à la spectrométrie de masse en arrivant au laboratoire, et j’ai toujours pu compter sur toi lorsque j’avais des questions ou des doutes. On s’est bien pris la tête sur ce traitement de données DIA, la normalisation des temps de rétention etc… Merci aussi pour ton humour aiguisé, c’était du haut niveau ! Je te souhaite le meilleur à Boston, et apparemment ça te plaît, que ça continue ! Merci Joanna, j’ai vraiment aimé travailler avec toi. Tu es intéressée, consciencieuse et rigoureuse, et je suis fier de voir que tu es arrivée aussi vite à ce niveau de maîtrise. Je suis sûr que la suite de ta thèse va très bien se passer. Merci également pour ton humour un peu décalé, et pour tous ces « désolé » qui nous ont valu de très bons gâteaux Merci pour ton aide et ton soutien notamment pour la fin de ma thèse, pour retraiter les données, terminer la mise en page du manuscrit... Bon courage pour la fin de ta thèse, et pour t’occuper de notre instrument préféré ! :3 Le bureau des gros… Merci infiniment pour ces 3 années et quelques. C’est comme si le destin avait rassemblé les plus gros morphales du labo dans le même bureau ! Merci pour la bonne humeur qui y régnait, toutes ces choses qu’on a mangées ensemble, tous ces moments qu’on a partagés, et tous ces chats ! Merci Nina, ma Chouquette, pour ton humour très décalé, pour ta gentillesse, les apéros, la colocation à San Antonio, etc. Je te souhaite le meilleur pour la suite, ainsi qu’à Martoune et Kevin Jean Bernard Edouard Raphaël. Merci Margaux, Margrosse, nos échanges de mots doux vont me manquer ! Merci pour ton soutien, et toutes les franches rigolades qu’on a eues. Nina et Margaux, j’ai presque cohabité avec vous pendant plus de 3 ans, rarement tôt le matin, mais des fois tard le soir, lorsque l’inspiration était à son paroxysme : c’est là qu’ont été créés le kidnappeur de tractopelle (et son jumeau encore plus maléfique le kidnappeur de kidnappeur de tractopelle), les blagues de photos de vacances, les canulars téléphoniques… Marianne et Aurélie, merci infiniment pour votre gentillesse, pour votre soutien… Vous avez toujours été là pour me faire rire, me nourrir (c’est vous les meilleures pâtissières ), m’héberger… Et aussi pour cette dernière soirée, vous étiez impliquées comme si c’était votre propre thèse, et on l’a finie ensemble! Merci pour tout, je n’y serais pas arrivé sans vous. Marianne, ma voisine de bureau, j’ai vraiment aimé travailler à côté de toi, te charrier… Je te souhaite vraiment le meilleur au Canada ainsi qu’à Fadi.
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