Structure/property studies of polymeric gene delivery using a library of poly(β-amino esters)  Daniel G. Anderson, Akin Akinc, Naushad Hossain, Robert Langer  Molecular Therapy  Volume 11, Issue 3, Pages 426-434 (March 2005) DOI: 10.1016/j.ymthe.2004.11.015 Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

Fig. 1 Acrylate and amino monomers used to synthesize poly(β-amino ester) library. Molecular Therapy 2005 11, 426-434DOI: (10.1016/j.ymthe.2004.11.015) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

Fig. 2 Molecular weight of polymer library. Mean polymer molecular weights organized from best transfecting (left) to worst (right) and amine/acrylate ratio from high (front) to low (back). Molecular weight was measured by GPC relative to polystyrene standards. Red arrows indicate polymers made with DMSO as described under Experimental Procedures. Molecular Therapy 2005 11, 426-434DOI: (10.1016/j.ymthe.2004.11.015) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

Fig. 3 Transfection proficiency of the polymer library. COS-7 cells were transfected with polymer/DNA complexes at six polymer/DNA ratios in quadruplicate: 10:1, 20:1, 30:1, 40:1, 60:1, and 100:1. The average transfection level in nanograms of luciferase per well of the optimal polymer/DNA ratio is presented. Polymers are organized from highest transfecting (C32) to lowest transfecting (II32) and amine/acrylate ratio from high (front) to low (back). Red arrows indicate polymers made with DMSO as described under Experimental Procedures. Molecular Therapy 2005 11, 426-434DOI: (10.1016/j.ymthe.2004.11.015) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

Fig. 4 Transfection proficiency relative to polymer molecular weight and polymer/DNA ratio. COS-7 cells were transfected with polymer/DNA complexes at six polymer/DNA ratios in quadruplicate: 10:1, 20:1, 30:1, 40:1, 60:1, and 100:1. The average transfection level in nanograms of luciferase per well is presented for all ratios as a function of molecular weight. Molecular Therapy 2005 11, 426-434DOI: (10.1016/j.ymthe.2004.11.015) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

Fig. 5 Relationship of polymer/DNA particle size and transfection efficiency. Polymer/DNA complexes were formed at polymer/DNA ratios optimal for transfection (Fig. 3). Data are organized from highest transfecting (left) to lowest transfecting (right) and amine/acrylate ratio from high (front) to low (back). Red arrows indicate polymers made with DMSO as described under Experimental Procedures. Molecular Therapy 2005 11, 426-434DOI: (10.1016/j.ymthe.2004.11.015) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions

Fig. 6 Relationship of polymer/DNA complex surface charge and transfection efficiency. Polymer/DNA complexes were formed at polymer/DNA ratios optimal for transfection. ζ potential is presented in mV. Data are organized from highest transfecting (left) to lowest transfecting (right) and amine/acrylate ratio from high (front) to low (back). Red arrows indicate polymers made with DMSO as described under Experimental Procedures. Molecular Therapy 2005 11, 426-434DOI: (10.1016/j.ymthe.2004.11.015) Copyright © 2004 The American Society of Gene Therapy Terms and Conditions