CHARACTERIZATION OF GENETIC DELETIONS IN BECKER MUSCULAR-DYSTROPHY USING MONOCLONAL-ANTIBODIES AGAINST A DELETION-PRONE REGION OF DYSTROPHIN

We have produced a new panel of 20 monoclonal antibodies (mAbs) agains t a region of the dystrophin protein corresponding to a deletion-prone region of the Duchenne muscular dystrophy gene (exons 45-50). We show that immuno-histochemistry or Western blotting with these ''exon-spec ific'' mAbs can provide a valuable addition to Southern blotting or PC R methods for the accurate identification of genetic deletions in Beck er muscular dystrophy patients. The antibodies were mapped to the foll owing exons: exon 45 (2 mAbs), exon 46 (6), exon 47 (1), exons 47/48 ( 4), exons 48-50 (6), and exon 50 (1), PCR amplification of single exon s or groups of exons was used both to produce specific dystrophin immu nogens and to map the mAbs obtained. PCR-mediated mutagenesis was also used to identify regions of dystrophin important for mAb binding. Bec ause the mAbs can be used to characterize the dystrophin produced by i ndividual muscle fibres, they will also be useful for studying ''rever tant'' fibres in Duchenne muscle and for monitoring the results of myo blast therapy trials in MD patients with deletions in this region of t he dystrophin gene. (C) 1995 Wiley-Liss, Inc.