At-line quantification of bioactive antibody in bioreactor by surface plasmon resonance using epitope detection.
We report an innovative at-line method to monitor concentration of bioactive antibody (i.e., antibody with conserved antigen-binding activity) secreted during bioreactor culture by the use of surface plasmon resonance (SPR)-based biosensor technology. In a first series of experiments, conditions for SPR-based measurements were validated off-line by monitoring bioactive antibody concentration in conditioned medium from 500-ml baffled flask hybridoma cell cultures. A fully automated experimental setup in which the SPR-based biosensor was harnessed to a bioreactor was then used at-line to monitor the concentration of bioactive antibody produced in a 3.5-L bioreactor. Quantitative SPR measurements performed both at-line and off-line were in excellent agreement with quantitative Western blotting followed by densitometry analyses. Thus, our experimental study confirms that SPR biosensors can be applied to at-line quantification of correctly folded proteins that are secreted by cells cultured in a bioreactor. Our experimental approach represents a novel and robust analytical strategy to be applied to the control and optimization of the production of bioactive secreted proteins.
Chavane
Natalie
N
Department of Chemical Engineering, Ecole Polytechnique de Montréal, Montréal, QC H3C3A7, Canada.
Jacquemart
Renaud
R
Hoemann
Caroline D
CD
Jolicoeur
Mario
M
De Crescenzo
Gregory
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D019149:Bioreactors; D015374:Biosensing Techniques; D015153:Blotting, Western; D002460:Cell Line; D003720:Densitometry; D000939:Epitopes / Q000032:analysis*; D006825:Hybridomas; D008024:Ligands; D008297:Male; D017430:Prostate-Specific Antigen / Q000378:metabolism; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance / Q000379:methods*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D008024:Ligands; D017430:Prostate-Specific Antigen