Animal Blood B Booster Kit

Dendritics routinely uses the hybridoma technology described by Köhler and Milstein, where the freshly isolated splenocytes of immunized mice, are fused with the Ig non secreting murine myeloma SP2/0 cell line to immortalize Ig secreting B lymphocytes. The resulting repertoire is generally limited either by the low immunogenicity of the antigen, or by the natural immunity of the mouse. Because the Blood B Booster system (DDXK-HuBBB) was demonstrated as being powerful for human B cells, we attempted to improve the efficiency of the hybridoma technology by introducing an activation step before the fusion in an attempt to give access to a broader repertoire. Briefly, after the immunization program, freshly isolated splenocytes were separately stimulated according to two stimulation systems based on B cell activation through CD40 triggering. We have assumed that the inter-species homology of the CD40 antigen will allow non human B cell activation by a monoclonal antibody raised against human CD40. After the demonstration that the introduction of the stimulation step before fusion significantly increased the clonal diversity of the immortalized murine B cell populations, this process was successfully extended to other animal species starting from peripheral blood samples (Razanajaona-Doll D. et al, AAI 98^th annual meeting, Immunology 2011™, San Francisco, CA, USA).
The kit AnBBB^® was packaged so as to make it very easy to handle for the scientists ; the kit AnBBB^® contains all the reagents needed to obtain around 3.10³ independent B cell clones from 6.10⁷ splenocytes or PBMC using six 96 wells culture plates for the stimulations and ten 96 wells plates for the fusion step. The resulting cells are available for a wide range of studies including genomic. transcriptomic and proteomic fields.