Toward in Vivo Imaging of Heart Disease Using a Radiolabeled Single-Chain Fv Fragment Targeting Tenascin-C

Antibodies specific to a particular target molecule can be used as analytical reagents, not only for in vitro immunoassays but also for noninvasive in vivo imaging, e.g., immunoscintigraphies. In the latter case, it is important to reduce the size of antibody molecules in order to achieve suitable in vivo “diagnostic kinetics” and generate higher-resolution images. For these purposes, single-chain Fv fragments (scFvs; Mr < 30 kDa) have greater potential than intact immunoglobulins (∼150 kDa) or Fab (or Fab′) fragments (∼50 kDa). Our recent observation of enhanced tenascin-C (Tnc) expression at sites of cardiac repair after myocardial infarction prompted us to develop a radiolabeled scFv against Tnc for in vivo imaging of heart disease. We cloned the genes encoding the heavy and light chain variable domains of the mouse anti-Tnc monoclonal antibody 4F10, and combined them to create a single gene. The resulting scFv-4F10 gene was expressed in E. coli cells to produce soluble scFv proteins. scFv-4F10 has an ...