title: Quantification of antibody production of individual hybridoma cells by surface plasmon resonance imaging.
abstract: Surface plasmon resonance imaging (SPRi) is most frequently used for the label-free measurement of biomolecular interactions. Here we explore the potential of SPRi to measure antibody production of individual hybridoma cells. As a model system, cells from a hybridoma, producing monoclonal antibodies recognizing epithelial cell adhesion molecule (EpCAM), were used. Recombinant human EpCAM protein was immobilized on an SPR sensor and hybridoma cells were introduced into an IBIS MX96 SPR imager and the SPRi response was followed for 10h. SPRi responses were detected on the spots of the sensor only where ligands of the produced antibody were present. By measuring the SPRi signals on individual cells the antibody production of the individual cells was measured and production rates were calculated. For 53 single EpCAM hybridoma cells the production ranged from 0.16 to 11.95 pg (mean 2.96p g per cell, SD 2.51) over a period of 10 h. Antibody excretion per cell per hour ranged from 0.02 to 1.19 pg (mean 0.30, SD 0.25). Here we demonstrate for the first time that antibody production of individual cells can be measured and quantified by SPRi, opening a new avenue for measuring excretion products of individual cells.
authors.affiliation: Medical Cell BioPhysics Group, MIRA Institute, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands. Electronic address: i.stojanovic@utwente.nl.
authors.forename: Ivan
authors.initials: I
authors.lastname: Stojanović
authors.affiliation: Medical Cell BioPhysics Group, MIRA Institute, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands; IBIS Technologies B.V., 7521 PR Enschede, The Netherlands.
authors.forename: Thomas J G
authors.initials: TJ
authors.lastname: van der Velden
authors.affiliation: Medical Cell BioPhysics Group, MIRA Institute, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands.
authors.forename: Heleen W
authors.initials: HW
authors.lastname: Mulder
authors.affiliation: Medical Cell BioPhysics Group, MIRA Institute, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands; IBIS Technologies B.V., 7521 PR Enschede, The Netherlands.
authors.forename: Richard B M
authors.initials: RB
authors.lastname: Schasfoort
authors.affiliation: Medical Cell BioPhysics Group, MIRA Institute, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands.
authors.forename: Leon W M M
authors.initials: LW
authors.lastname: Terstappen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism*; D000951:Antigens, Neoplasm / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000276:immunology; D000071858:Epithelial Cell Adhesion Molecule; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D000906:Antibodies; D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule
keywords: Antibody production; EpCAM; Hybridoma; SPR cytometry; SPRi