The objective of this project is to generate a library of well-characterized, antihuman skeletal myosin monoclonal antibodies to be used as sensitive and specific reagents to ascertain and characterize the diversity of such isoforms and investigate their modification under physiologic and pathologic conditions. The McAbs will be generated using human skeletal myosin light chains as immunogen to BALB/C mice. Their specificity will be characterized as to species specificity, heavy versus light chain specificity, subfragment and macromolecular location of recognized epitope, as well as to native isozyme specificity. This will be done with the use of one and two dimensional electrophoresis as well as electrophoresis of myosin under nondissociating conditions coupled with Western blotting to which McAbs will be reacted and displayed using a second radioactively labeled antibody and developed autoradiographically. Macromolecular epitope localization will be done with immuno-electron microscopy. In addition, the myosin isoform-specific McAbs generated, will be used in immuno-affinity chromatography to isolate and purify human skeletal myosin isoenzymes for use in future work aimed at correlating the heterogeneity of human myosins with functional properties and ultimately with clinical manifestations of neuromuscular disease. As these McAbs are generated and characterized, they will begin to be applied to survey the normal diversity of myosin isoforms in human skeletal muscle and also used to examine pathological specimens for abberations or abnormalities in the distribution or structure of myosin in neuromuscular disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Clinical Investigator Award (CIA) (K08)
Project #
1K08NS001123-01
Application #
3083841
Study Section
Neurological Disorders Program Project Review B Committee (NSPB)
Project Start
1986-04-01
Project End
1991-03-31
Budget Start
1986-04-01
Budget End
1987-03-31
Support Year
1
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Suny Downstate Medical Center
Department
Type
Schools of Medicine
DUNS #
068552207
City
Brooklyn
State
NY
Country
United States
Zip Code
11203