Production, characterization and application of poly(β-amino ester) based nanoparticular gene carrier formulations synthesized by different methods

In gene transfer, vectors are used to carry genetic material Vectors should have low toxic effects and should selectively and efficiently carry genetic material into the cell. Vectors are divided into two groups as viral and non-viral vectors. In viral vectors, genetic material is carried by viruses. In non-viral vectors, genetic material is carried by natural and synthetic materials. Viral vectors are quite effective in terms of efficiency of reaching the target cell. However, their applications are limited in terms of production difficulties and reliability. Due to these disadvantages, there has been an increase in the tendency towards non-viral vectors that are easy to produce, have low toxic effects, high loading capacity and low immune response. The vector to be used in gene transfer should be target cell selectivity, metabolically degradable, resistant to the immune system, have the least toxic effect and be able to maintain the genetic material at a certain level for as long as needed.Within the scope of this thesis, synthesis reactions of PBAE, a non-viral vector, will be carried out by conventional method, microwave method and catalyst method. Characterization and optimization studies of polymer formulations obtained by 3 different synthesis methods will be carried out. Nanoparticulate systems will be obtained by nanoprecipitation method. The zeta potentials, nanoparticle sizes and polydispersity indices of the obtained nanoparticulate systems will be determined and optimized. According to the results of structure elucidation and optimization studies of nPBAEs, complexes and ionic interactions of matching nPBAEs with plasmid DNA will be examined. pDNA binding properties of nPBAEs will be analyzed in gel electrophoresis device. In vitro transfection efficiency and cytotoxicity will be analyzed. As a result of these studies, for PBAEA3 (1.1:1 diamine:diacrylate molar ratio) synthesized by the conventional method, the nanoparticle size was found to be 107.1±3.2, the PDI value was 0.142±0.022 and the zeta potential was 13.4±4.3 for the nanoparticle system synthesized at the 1200:300 deionized water:polymer ratio and for PBAEA4 (1.5:1 diamine:diacrylate molar ratio), the nanoparticle size was found to be 106.5±5.5, the PDI value was 0.213±0.013 and the zeta potential was 10.7±3.2 for the nanoparticle system synthesized at the 1200:300 deionized water:polymer ratio. The nanoparticle systems obtained at these ratios showed successful transfection efficiency.