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

Polymers (PONI) were synthesized by ring-opening metathesis polymerization using third generation Grubbs' catalyst as previously described. 65 In brief, solutions in dichloromethane of guanidium functionalized monomer and Grubbs' catalyst were placed under freeze thawing cycles for degassing. After warming the solutions to room temperature, the degassed monomer solutions were administrated to degassed catalyst solutions and allowed to stir for 30 minutes. The polymerization reaction was terminated by the addition of excess ethyl vinyl ether. The reaction mixture was further stirred for another 30 min. The resultant polymers were precipitated from excess hexane or diethyl ether anhydrous, filtered, washed and dried under vacuum to yield a light-yellow powder. polymers were characterized by 1 H NMR and gel permeation chromatography (GPC) to assess chemical compositions and molecular weight distributions, respectively. Subsequent to deprotection of Boc functionalities, polymer was dissolved in the DCM with the addition of TFA at 1:1 ratio. The reaction was allowed to stir for 4 hours and dried under vacuum. Excess TFA was removed by azeotropic distillation with methanol. Afterwards, the resultant polymers were re-dissolved in DCM and precipitated in anhydrous diethyl ether, filtered, washed and dried. Polymers were then dissolved in water and transferred to Biotech CE dialysis tubing membranes with a 3000 g/mol cutoff and dialyzed against RO water (2−3 days). The polymers were then lyophilized dried to yield a light white powder.