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Anal Biochem. 2015 Sep 15;485:112-8. doi: 10.1016/j.ab.2015.06.018. Epub 2015 Jun 18.

Quantification of antibody production of individual hybridoma cells by surface plasmon resonance imaging.

Author information

1
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.
2
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.
3
Medical Cell BioPhysics Group, MIRA Institute, Faculty of Science and Technology, University of Twente, 7500 AE Enschede, The Netherlands.

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.

KEYWORDS:

Antibody production; EpCAM; Hybridoma; SPR cytometry; SPRi

PMID:
26095397
DOI:
10.1016/j.ab.2015.06.018
[Indexed for MEDLINE]

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