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

. CC-BY-NC-ND 4.0 International license author/funder. It is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.15.037564 doi: bioRxiv preprint 150 nm carboxylate nanoparticles (Phosphorex) were exchanged into 50 mM MES buffer at pH 5.2 via gel filtration column. N-Hydroxysulfosuccinimide (sulfo-NHS) was added to the particles at 0.275 mg/mL, prior to incubation for 3 minutes at room temperature. EDCI was then added to the particles at 0.1 mg/mL, prior to incubation for 15 minutes at room temperature. IgG was added to the particle mixture at 200 IgG per nanoparticle, prior to incubation for 3 hours at room temperature while vortexing. For radiotracing, 125 I-labeled IgG was added to the reaction at 5% of total IgG mass. The IgG/particle mixture was diluted with 10-fold volume excess of pH 5.2 MES buffer and the diluted mixture was centrifuged at 12000xg for 3 minutes. Supernatant was discarded and PBS with 0.05% BSA was added at desired volume before resuspending the particles via sonication probe sonication (three pulses, 30% amplitude). Particle size was assessed via DLS after resuspension, and particles were used immediately after DLS assessment.