|Design and Commercial Production of Mouse Hybridomas to Produce Antibody to FMRP
by Ivan Jeanne Weiler , 1/1/2005
Antibodies to specific proteins, such as FMRP, are currently the most important tools we have to study where the protein goes and what things it interacts with in brain cells. The best antibodies are "monoclonal", because these cells reproduce indefinitely and will continue to produce this specific antibody. (Antibodies form the basis of the body’s immune system – they recognize and grab onto foreign proteins, viruses, etc. that may pose a threat.)
Most Fragile X researchers still use a monoclonal antibody (1C3) which robustly recognizes FMRP. However, there are problems with it. First, it reacts slightly with another protein, FXR1p, as well as FMRP, so that if we use it to stain tissue, we cannot be sure we are staining only FMRP. This would be important, for example, when determining whether gene therapy had succeeded in helping cells to produce FMRP.
Much of the current research depends on the ability to purify protein clusters which contain FMRP with associated RNA and other proteins, using a technique called immunoprecipitation. For reasons we do not understand, 1C3 fails to precipitate FMRP under normal conditions. Other laboratories have made antibodies to FMRP which will immunoprecipitate, but cannot be used in staining. Our aim is to produce an antibody which will do both.
Because commercial labs have concentrated on developing tricks to elicit monoclonal antibodies with more success than the average university lab, we are contracting with Strategic Biosolutions to produce new monoclonals. We have identified three promising sequences in the FMRP molecule which have not been used before. The company will produce candidate clones based on these sequences and send the clones to us for selection of the best candidates.
If we succeed in obtaining our "dream antibody" we will donate the cells to the Iowa antibody resource which will make the line available to the entire research community.