Abstract: Methods and compositions for the production of phospholipid-ligand bioconjugates and uniform targeted microbubbles are provided. These methods and compositions find use in ultrasound and molecular imaging applications related to cancer and other diseases. The methods of the present disclosure comprise contacting a phospholipid comprising a maleimide containing functional group with a ligand comprising a C terminal cysteine residue. The methods disclosed herein solves the problems in producing ready-to-use and clinically translatable ultrasound molecular imaging agents by incorporating small protein ligands engineered to bind against biomarkers representing pathological angiogenesis or abnormal cells. The methods also overcome the current limitations in producing uniformly targeted microbubbles in a scalable, economical and reproducible manner.