Fig. 1. Cultured Hybridoma Cells after Fusion. The cells are shown under 100 x magnification.
To develop high quality monoclonal antibodies and cell lines, our technicians follow a distinctive procedure designed to maximize your chance of success.
Fusion
We harvest B-cells not only from the spleen (as others do), but also from multiple lymph nodes. To ensure the successful generation of hybridoma, we would perform 3 parallel fusions for cells harvested from each mouse.
Selection of Clones for Subcloning
We would pick a large number of initial clones (~360) for each peptide antigen, about one quarter of which (~90) would turned out to be positive by ELISA test. A second round of ELISA tests is performed, in which the concentration of each peptide antigen undergoes serial dilutions until no signal can be detected. Clones are ranked by relative affinity (greater maximum dilution factor = greater affinity). 10 clones with highest ranking are further chosen for sub cloning.