TY  - JOUR
A1  - Kuhne, Maren
A1  - Dippong, Martin
A1  - Flemig, Sabine
A1  - Hoffmann, Katrin
A1  - Petsch, Kristin
A1  - Schenk, Jörg A.
A1  - Kunte, Hans-Jörg
A1  - Schneider, Rudolf J.
T1  - Comparative characterization of mAb producing hapten-specific hybridoma cells by flow cytometric analysis and ELISA
JF  - Journal of immunological methods
N2  - A novel method that optimizes the screening for antibody-secreting hapten-specific hybridoma cells by using flow cytometry is described. Cell clones specific for five different haptens were analyzed. We selectively double stained and analyzed fixed hybridoma cells with fluorophore-labeled haptens to demonstrate the target-selectivity, and with a fluorophore-labeled anti-mouse IgG antibody to characterize the level of surface expression of membrane-bound IgGs. ELISA measurements with the supernatants of the individual hybridoma clones revealed that antibodies from those cells, which showed the highest fluorescence intensities in the flow cytometric analysis, also displayed the highest affinities for the target antigens. The fluorescence intensity of antibody-producing cells corresponded well with the produced antibodies' affinities toward their respective antigens. Immunohistochemical staining verified the successful double labeling of the cells. Our method makes it possible to perform a high-throughput screening for hybridoma cells, which have both an adequate IgG production rate and a high target affinity. (C) 2014 Elsevier B.V. All rights reserved.
KW  - Immunization
KW  - Hapten
KW  - Monoclonal antibodies
KW  - Hybridoma
KW  - Flow cytometry
KW  - ELISA
Y1  - 2014
U6  - https://doi.org/10.1016/j.jim.2014.07.004
SN  - 0022-1759
SN  - 1872-7905
VL  - 413
SP  - 45
EP  - 56
PB  - Elsevier
CY  - Amsterdam
ER  -