Quantitative estimation of dystrophin protein: a sensitive and convenient "two-antibody sandwich" ELISA.
As dystrophin protein, the protein product of Duchenne muscular dystrophy (DMD) gene, represents only 0.002 approximately 0.03% of the total muscle proteins and human dystrophin protein has not been purified, quantitative estimation of this protein has been difficult. We describe a sensitive, reliable and convenient "two-antibody sandwich" enzyme-linked immunosorbent assay (ELISA) using commercially available monoclonal antibodies. This system, using a capture antibody specific for carboxyl terminus and two different detection antibodies for the mid-rod domain and the amino-terminal domain, is highly specific for dystrophin, since muscle specimens from DMD patients gave almost zero response (n = 3, 0.38 approximately 0.45%; expressed as a percentage of normal muscle tissue). This assay should prove to be an accessible and useful tool for the diagnosis of DMD/BMD and for the evaluation in clinical trials such as myoblast transfer and gene therapy.
Ishikawa
Y
Y
Department of Pediatrics, National Sanatorium Yakumo Hospital, Japan.
Ishikawa
Y
Y
Minami
R
R
D000911:Antibodies, Monoclonal; D016189:Dystrophin / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D009132:Muscles / Q000378:metabolism; D009136:Muscular Dystrophies / Q000378:metabolism; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D016189:Dystrophin