Radiolabelled monoclonal antibodies against human cardiac myosin.
Myocardial infarction (MI) can be monitored using several protein markers including human cardiac myosin (HCM). Monoclonal antibodies were raised against HCM by hybridoma technique. Antimyosin antibody producing clones were identified by ELISA and monoclonality was established by limiting dilution. The antibodies were purified, isotyped and their cross reactions with myosin from other species were estimated. All the clones showed negligible cross reaction with rabbit myosin, but reacted with bovine skeletal myosin to different extents (40-100%). The most avid antibody Mab 4G4 which also strongly reacted with rat cardiac myosin, was labelled with 125I using different oxidising agents such as iodogen, chloramine-T and lactoperoxidase. More than 95% pure radiolabelled antibody could be obtained by gel filtration. The immunoreactivity was retained. Mab 4G4 was also labelled with 99mTc using stannous tartrate as the reducing agent. Radiolabelling yield was approximately 60%, the purity was >95%. Both the radiolabelled preparations were tested for biodistribution in rats--both normal and those with induced MI. Approximately 0.7 % of the injected activity/g was found in the infarcted region and the accumulation of activity in the infarcted heart was 1.5 times that in the normal heart. A very high percentage of activity (80%) accumulated in the thyroid. With further optimisation of labelling and use of F(ab')2 fragments, better delineation of the infarct sites may become possible.
Bapat
K N
KN
Radiopharmaceutical Division, Bhabha Atomic Research Centre, Mumbai 400 085, India.
Venkatesh
M
M
Sarma
H D
HD
Pillai
M R
MR
Sainis
K B
KB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D024722:Cardiac Myosins / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D011868:Radioisotopes
D000911:Antibodies, Monoclonal; D011868:Radioisotopes; D024722:Cardiac Myosins