1. Construction of dystrophin fusion proteins to raise targeted antibodies to different epitopes
- Author
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Antoon F.M. Moorman, H.B.M. van Paassen, J.T. den Dunnen, G.J.B. van Ommen, Glenn E. Morris, H.B. Ginjaar, Nguyen thi Man, and Other departments
- Subjects
musculoskeletal diseases ,medicine.drug_class ,Recombinant Fusion Proteins ,Duchenne muscular dystrophy ,Restriction Mapping ,Biophysics ,Monoclonal antibody ,Biochemistry ,Epitope ,Dystrophin ,Epitopes ,Antibody Specificity ,Structural Biology ,Duchenne muscular dystrophy: P20 mutation hotspot ,Genetics ,medicine ,Humans ,Molecular Biology ,Peptide sequence ,biology ,Muscles ,Nucleic acid sequence ,Cell Biology ,medicine.disease ,Immunohistochemistry ,Molecular biology ,Fusion protein ,Polyclenal antiserum ,pATH2 fusion protein ,Polyclonal antibodies ,Antibody Formation ,biology.protein ,pEX1 fusion protein - Abstract
For the study of the structure and function relationship of dystrophin, defective in DMD, and for diagnostic purposes it is important to dispose of untibodies against different parts of the protein. We have made five different constructs for the expression of fusion proteins containing parts of the four domains of dystrophin. Two different recombinant expression vectors, pATH2 and pEX1, were used. Rabbits were immunized with the fusion products and several polyclonal antibodies were raised. At a later stage, monoclonal antibodies were also raised to some of the fusion proteins. One polyclonal antibody, named P20 AB, is directed against the region covering amino acid sequence 1749–2248 or the nucleotide sequence 5456–6953 of the mRNA, which corresponds to the major deletion-prone region of the DMD gene. We show the particular value, sensitivity and specificity of the P20 AB in dystrophin analysis.
- Published
- 1992