Nanoparticles  act  as  a  promising  system  for  targeted  delivery  of  drugs  and  as  an  effective  route  of  drug  administration.  In  this  study,  polysorbate  80  coated  nanoparticles  of  serratiopeptidase  were  formulated  and  aimed for  the  treatment  of  blood  clots  in  brain.  Serratiopeptidase exerts effective activity against blood clotting and has ability to dissolve non-living tissues,   blood clots, cysts, atherosclerotic clots. Different  nanoparticle  formulations  of  serratiopeptidase  were  prepared  with  different  concentrations  of  chitosan  and  tripolyphosphate  using  ionic  gelation  method.  The  nanoparticles were coated using polysorbate 80 and were characterized  and  evaluated  for  different  parameters  such  as  particle  size,  entrapment  efficiency,  zeta  potential and  transmission  electron  microscopy.  The  in  vitro  drug  release  of  prepared  nanoparticles  was  studied  in  phosphate  buffer  (pH 7.4).  The  results  indicated  that  the  developed  nanoparticle  formulation  could  be  established  as  a  promising carrier  for  active  targeting  into  brain  to  dissolve  blood  clots.