Fig. 1.

Optimization of F(ab’)2 conjugation to polymeric nanoparticles. a Scheme of antibody fragment conjugation to the surface of pre-formulated maleimide-functionalized PEG-PLGA polymeric nanoparticles (NPs). b A non-reducing SDS–PAGE gel stained with Coomassie Brilliant Blue is shown following IdeS-mediated cleavage of anti-CD8a and rat IgG2b isotype control antibodies. c A Western blot following reducing SDS–PAGE of CD8a-targeting nanoparticles developed with Fab-specific (left panel) or Fc-specific antibodies (right panel); lane 1: uncoated nanoparticles, lane 2: nanoparticles without antibody reduction before conjugation, lane 3: anti-CD8a nanoparticles with the antibody reduced using 0.5 mM DTT before conjugation, lane 4: anti-CD8a F(ab’)2 and Fc cleavage product as a positive control. d Scanning electron microscopy images of blank NP formulations before and after F(ab’)2 conjugation; scale bar = 500 nm