CONFIDENTIAL, NOT FOR DISCLOSURE This is confidential

By Canelle Quentin
Dissertation submitted to the Faculty of Sciences of the Université Libre de Bruxelles,
in partial fulfillment of the requirements for the degree of
Doctor in agronomy and bioengineering
2015
Jury Member:
Professor Erik Goormaghtigh, Président
Professor Michel Vandenbranden, Secrétaire
Professor Fabrice Homblé, Promoteur
phD Pol Harvengt
phD Robbert Van Der Most
CONFIDENTIAL, NOT FOR DISCLOSURE
This is confidential information of GSK Vaccines. This document is only for review and should
not be shared further.
Acknowledgment
Avant de vous proposer la lecture de cette dissertation, il me tenait à coeur de remercier les
différentes personnes qui ont contribué à la réalisation de celle-ci.
Ce travail n’aurait pu être réalisé sans l’aide du Professeur Fabrice Homblé qui a pris en
charge l’encadrement académique ni sans l’aide du Dr Pol Harvengt qui a permis mon
insertion au sein GSK vaccines.
Ce travail n’aurait pu se faire sans la société GSK vaccines qui a pris en charge le financement
de la recherche. Je tiens à remercier tout particulièrement le département Vaccine Research
dans son intégralité et plus particulièrement Marcelle Van Mechelen, Stéphane
Temmerman, Sandrine Tonon, Sofia Buonocore, Philippe Auquier, Nabila Amanchar, Hajar
Larbi, Stéphanie Quique pour l’accueil chaleureux qui m’a été réservé au sein du groupe.
I also would like to warmly thank Robbert Van Der Most, Wivine Burny, Vincent Dewar, John
Barlow and Benoit Collignon for the science discussions we had and their motivation.
Je désire également exprimer ma gratitude envers le Dr Yvon Corazza pour son initiation à la
technologie BIAcore ainsi que pour sa créativité qu’il m’a permis de partager.
Je souhaite également remercier les membres du jury qui prêteront attention à ce manuscrit
lors de leur lecture avisée.
Pour leur présence ou leur contribution variée mais néanmoins indispensable, je tiens
également à rendre hommage à Muriel Feyssaguet et son équipe, Thierry Ollinger, Michaël
Dehottay, Karine Gillard, Philippe Jehoulet, Geneviève Jadot, Sylvie Collignon, Géraldine
Minet, Pilar Garcia, Bernard hoyois, Benoit Vincart, Cédric Vanderhaegen, Aurélie Chalon,
Catherine Collignon, Carsten Walter, Caroline Hervé, Christel Verachtert, Christine Dantinne,
Françoise Villée, Jennifer Dalcq, Laurie Dumont, Loïc Le Gourrierec, Margherita Coccia,
Richard Ruez, Sara Lemos, Christine Van Eyck, Maryline Bauthier, Viviane Bechtold, Robert
Van Den Berg, Abdelatif Elouahabi, Sandra Morel, Alessandra Ferraro, et l’équipe de
Formulation.
D’un point de vue personnel, je tiens également à remercier mes proches et tout d’abord
mes parents qui ont tout mis en œuvre pour m’aider à réussir mes entreprises, quelles
qu’elles aient été ; ma compagne qui m’a supporté lors des moments de stress, de malheur
mais aussi de bonheur; mon frère et ma sœur qui sont des soutiens constants et
indéfectibles ; mes grands–parents qui m’ont toujours encouragé à leur manière ; mes amis
qui auront toujours pris la peine de s’inquiéter pour moi et de prendre de mes nouvelles.
À tous, encore une fois, merci.
Table of content
GENERAL INTRODUCTION ________________________________________ ERROR! BOOKMARK NOT DEFINED.
BIOMOLECULAR INTERACTION ____________________________________________ERROR! BOOKMARK NOT DEFINED.
Avidity and affinity ______________________________________________ Error! Bookmark not defined.
Kinetics _______________________________________________________ Error! Bookmark not defined.
Binding interaction assessment ____________________________________ Error! Bookmark not defined.
ANTIBODY AND HUMORAL RESPONSE ________________________________________ERROR! BOOKMARK NOT DEFINED.
Humoral response and avidity _____________________________________ Error! Bookmark not defined.
Immunological implication of avidity ________________________________ Error! Bookmark not defined.
LABEL FREE TECHNOLOGY IN SERUM ANALYSIS __________________________________ERROR! BOOKMARK NOT DEFINED.
Surface Plasmon Resonance (SPR) biosensors _________________________ Error! Bookmark not defined.
Non Specific binding _____________________________________________ Error! Bookmark not defined.
Interaction models ______________________________________________ Error! Bookmark not defined.
POLYCLONAL ANTIBODY INTERACTION ANALYSIS BY REAL TIME BIOSENSORS ______________ERROR! BOOKMARK NOT DEFINED.
PROPOSAL AND AIM OF THE RESEARCH _______________________________________ERROR! BOOKMARK NOT DEFINED.
Application 1 ___________________________________________________ Error! Bookmark not defined.
Application 2 ___________________________________________________ Error! Bookmark not defined.
D-Pan and Q-Pan application ______________________________________ Error! Bookmark not defined.
MATERIAL AND METHOD _________________________________________ ERROR! BOOKMARK NOT DEFINED.
SERUM SAMPLES _____________________________________________________ERROR! BOOKMARK NOT DEFINED.
SAMPLE TREATMENT ___________________________________________________ERROR! BOOKMARK NOT DEFINED.
ANTIGEN LIGANDS ____________________________________________________ERROR! BOOKMARK NOT DEFINED.
ANTIGEN IMMOBILIZATION_______________________________________________ERROR! BOOKMARK NOT DEFINED.
DECONVOLUTION STRATEGY ______________________________________________ERROR! BOOKMARK NOT DEFINED.
Monoclonal Antibody selection ____________________________________ Error! Bookmark not defined.
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This is confidential information of GSK Vaccines. This document is only for review and should
not be shared further.
Additivity verification ____________________________________________ Error! Bookmark not defined.
NON SPECIFIC BINDING OPTIMIZATION _______________________________________ERROR! BOOKMARK NOT DEFINED.
APPLICATION 1 EXPERIMENT _____________________________________________ERROR! BOOKMARK NOT DEFINED.
APPLICATION 2 EXPERIMENT _____________________________________________ERROR! BOOKMARK NOT DEFINED.
D-PAN AND Q-PAN EXPERIMENT __________________________________________ERROR! BOOKMARK NOT DEFINED.
RESULTS _______________________________________________________ ERROR! BOOKMARK NOT DEFINED.
DECONVOLUTION STRATEGY ______________________________________________ERROR! BOOKMARK NOT DEFINED.
Monoclonal antibody selection ____________________________________ Error! Bookmark not defined.
Additivity verification ____________________________________________ Error! Bookmark not defined.
Discussion _____________________________________________________ Error! Bookmark not defined.
NON SPECIFIC BINDING _________________________________________________ERROR! BOOKMARK NOT DEFINED.
Discussion _____________________________________________________ Error! Bookmark not defined.
APPLICATION 1 ______________________________________________________ERROR! BOOKMARK NOT DEFINED.
Specific antibody content _________________________________________ Error! Bookmark not defined.
BIAcore® data’s _________________________________________________ Error! Bookmark not defined.
Association ____________________________________________________ Error! Bookmark not defined.
Dissociation ____________________________________________________ Error! Bookmark not defined.
Statistical analysis_______________________________________________ Error! Bookmark not defined.
Discussion _____________________________________________________ Error! Bookmark not defined.
APPLICATION 2 ______________________________________________________ERROR! BOOKMARK NOT DEFINED.
Specific antibody content _________________________________________ Error! Bookmark not defined.
Immobilization _________________________________________________ Error! Bookmark not defined.
BIAcore® Data’s ________________________________________________ Error! Bookmark not defined.
Binding association ______________________________________________ Error! Bookmark not defined.
Dissociation ____________________________________________________ Error! Bookmark not defined.
Discussion _____________________________________________________ Error! Bookmark not defined.
APPLICATION 3: D-PAN AND Q-PAN ________________________________________ERROR! BOOKMARK NOT DEFINED.
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CONFIDENTIAL, NOT FOR DISCLOSURE
This is confidential information of GSK Vaccines. This document is only for review and should
not be shared further.
HI titers _______________________________________________________ Error! Bookmark not defined.
Measurement of avidity by SPR (BIAcore®) analysis ____________________ Error! Bookmark not defined.
Association ____________________________________________________ Error! Bookmark not defined.
Dissociation ____________________________________________________ Error! Bookmark not defined.
Statistical analysis_______________________________________________ Error! Bookmark not defined.
Discussion _____________________________________________________ Error! Bookmark not defined.
CONCLUSIONS __________________________________________________ ERROR! BOOKMARK NOT DEFINED.
PERSPECTIVES __________________________________________________ ERROR! BOOKMARK NOT DEFINED.
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not be shared further.
ANNEXES ______________________________________________________ ERROR! BOOKMARK NOT DEFINED.
ANNEX 1: MATHEMATICAL TRANSFORMATION OF THE COMPLEX CONCENTRATION AT EQUILIBRIUMERROR!
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NOT
DEFINED.
ANNEX 2: FITTING OF THE MABS AND THE MAB MIXTURE __________________________ERROR! BOOKMARK NOT DEFINED.
ANNEX 3 : CORRELATION OF END ASSOCIATION LEVEL AT THE 2 CONCENTRATIONS _________ERROR! BOOKMARK NOT DEFINED.
ANNEX 4: CORRELATION BETWEEN SIMULATED STEADY STATES AT CONCENTRATION 0.05 AND 0.2 IUERROR! BOOKMARK
NOT
DEFINED.
ANNEX: 5 CORRELATION BETWEEN END ASSOCIATION LEVEL AND SIMULATED STEADY STATE ___ERROR! BOOKMARK NOT DEFINED.
ANNEX 6: % OF DISSOCIATION FOR ALL THE APPLICATION 1 SAMPLES AT 0.05 AND 0.2 CONCENTRATIONERROR! BOOKMARK NOT
DEFINED.
ANNEX 7: CORRELATION BETWEEN KD GLOBAL FITTING AND % OF DISSOCIATION ___________ERROR! BOOKMARK NOT DEFINED.
ANNEX 8: RELATION BETWEEN SAMPLE DILUTION AND END ASSOCIATION LEVEL FOR APPLICATION 1ERROR!
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DEFINED.
ANNEX 9: EXAMPLE OF BINDING CURVE (EXPERIMENTAL + COMPUTED FITTING) AT 0.2 CONCENTRATIONERROR! BOOKMARK NOT
DEFINED.
ANNEX 10: BINDING CHART OF APPLICATION 1 _________________________________ERROR! BOOKMARK NOT DEFINED.
ANNEX 11: END ASSOCIATION APPLICATION 2__________________________________ERROR! BOOKMARK NOT DEFINED.
ANNEX 12: END ASSOCIATION CORRECTION APPLICATION 2 ________________________ERROR! BOOKMARK NOT DEFINED.
ANNEX 13: APPLICATION 2 END ASSOCIATION LEVEL AND % OF DISSOCIATION ____________ERROR! BOOKMARK NOT DEFINED.
ANNEX 14: DISSOCIATION % FOR BOTH SURFACE AND SERUM SAMPLE _________________ERROR! BOOKMARK NOT DEFINED.
ANNEX 15: APPLICATION 2, % DISSOCIATION __________________________________ERROR! BOOKMARK NOT DEFINED.
ANNEX 16: DISSOCIATION CURVATURE_______________________________________ERROR! BOOKMARK NOT DEFINED.
BIBLIOGRAPHY _________________________________________________ ERROR! BOOKMARK NOT DEFINED.
iv
CONFIDENTIAL, NOT FOR DISCLOSURE
This is confidential information of GSK Vaccines. This document is only for review and should
not be shared further.