Hybridoma screening problem - (Jun/08/2001 )

We use a commercial hybridoma service to obtain monoclonal antibodies for haptens. To our surprise we often get no hybridoma cells which secrete antibodies which are similar to the antibodies in the mouse serum. Positive clones are very rare. We heard that with poorly soluble antigens the respective cells are in the lymph nodes and not in the spleen and therefore cannot be found as hybridoma. Has anyone experience with similar problems?

Michael, haptens are often too small to elicit a good immune response. Karl Landsteiner, in the 1920's proved that conjugating a nonimmunogenic homopolymer (carrier molecule) to your hapten will elicit a much better response. Increasing the size of your molecule will, to a point, increase its ability to be phagocytosed, making it a better immunogen. Solubility is key to immunizing the animal and getting the antigen to the spleen. BSA is a good carrier molecule that is also very soluble. You will just have to screen your positive clones against BSA later on to find specific antibodies. Hope that helps.