Supramolecular Organization Predicts Protein Nanoparticle Delivery to Neutrophils for Acute Lung Inflammation Diagnosis and Treatment

Jacob W. Myerson; Priyal N. Patel; Nahal Habibi; Landis R. Walsh; Yi-Wei Lee; David C. Luther; Laura T. Ferguson; Michael H. Zaleski; Marco E. Zamora; Oscar A. Marcos-Contreras; Patrick M. Glassman; Ian Johnston; Elizabeth D. Hood; Tea Shuvaeva; Jason V. Gregory; Raisa Y. Kiseleva; Jia Nong; Kathryn M. Rubey; Colin F. Greineder; Samir Mitragotri; George S. Worthen; Vincent M. Rotello; Joerg Lahann; Vladimir R. Muzykantov; Jacob S. Brenner

Lysozyme-dextran nanogels, crosslinked protein nanoparticles, E. coli, or adenovirus were similarly iodinated. At least 100 µL of particle suspension was added to a borosilicate tube containing two Iodobeads, prior to addition of 100 µCi of Na 125 I per 100 µL of suspension. Particles were incubated with radioiodine and Iodobeads for 30 minutes at room temperature, with gentle shaking every 10 minutes. To remove free iodine, particle suspensions were moved to a centrifuge tube, diluted in ~1 mL of buffer and centrifuged to pellet the particles (16000xg/30 minutes for nanogels, 16000xg/30 minutes for crosslinked protein particles, 10000xg/30 minutes for adenovirus, and 1000xg/10 minutes for E. coli). Supernatant was removed and wash/centrifugation cycles were repeated to assure >95% of radioactivity was associated with particles. Particles were resuspended by probe sonication (three pulses, 30% amplitude) for nanogels or crosslinked protein nanoparticles or pipetting for adenovirus or E. coli.