Effect of size and serum proteins on transfection efficiency of poly ((2-dimethylamino)ethyl methacrylate)-plasmid nanoparticles

• Published in: Pharmaceutical research 1996-07, Vol.13 (7), p.1038-1042
• Main Authors: CHERNG, J.-Y, VAN DE WETERING, P, TALSMA, H, CROMMELIN, D. J. A, HENNINK, W. E
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Biotechnology; Blood Proteins - metabolism; Cell Survival - drug effects; COS Cells - metabolism; Electricity; Fundamental and applied biological sciences. Psychology; Gene Expression - drug effects; Genetic engineering; Genetic technics; Macromolecular Substances; Methods. Procedures. Technologies; Particle Size; Plasmids - genetics; Plasmids - metabolism; Polymethacrylic Acids - metabolism; Polymethacrylic Acids - toxicity; Transfection; Vectors (cloning, transfer, expression). Insertion sequences and transposons
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1023/A:1016054623543

The aim of this study was to gain insight into the relation between the physical characteristics of particles formed by a plasmid and a synthetic cationic polymer (poly(2-dimethylamino)ethyl methacrylate, PDMAEMA) and their transfection efficiency. The PDMAEMA-plasmid particles were characterized by dynamic light scattering (size) and electrophoretic mobility measurements (charge). The transfection efficiency was evaluated in cell culture (COS-7 cells) using a pCMV-lacZ plasmid coding for beta-galactosidase as a reporter gene. It was shown that the optimal transfection efficiency was found at a PDMAEMA-plasmid ratio of 3 (w/w), yielding stable and rather homogeneous particles (diameter 0.15 micron) with a narrow size distribution and a slightly positive charge. Particles prepared at lower weight ratios, showed a reduced transfection efficiency and were unstable in time as demonstrated by DLS measurements. Like other cationic polymers, PDMAEMA is slightly cytotoxic. This activity was partially masked by complexing the polymer with DNA. Interestingly, the transfection efficiency of the particles was not affected by the presence of serum proteins. PDMAEMA is an interesting vector for the design of in vivo and ex vivo gene transfection systems.