The efficiency of human serum albumin on cationic polymer / DNA complex for gene delivery

• Main Authors: Yueh-Wan Luo, 羅月婉
• Other Authors: Jun-Lon Chen
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/71905284790835283057

碩士 === 國防醫學院 === 藥學研究所 === 97 === In order to improve the transfect efficiency of non-viral vector, cationic polymers were studied. Cationic polymer, Branched polyethylenimine ( B-PEI ), has been approved to be an efficient agent for gene delivery. PEI is a highly positive-charged organic macromolecule whose amino group condensed DNA and enhanced gene delivery into the nucleus of mammalian cells. Some reports demonstrated that the optimal molecular weight of PEI is in the range from 20 to 30 kDa to delivery DNA. But, Branched PEI also has cytotoxicity. Albumin is the most abundant plasma protein. Because of its biodegradability and lack of toxicity and immunogenicity, albumin is used as a drug carrier. Some reports demonstrated that HSA enhances DNA transfection by combining in lipoplexes. To decrease the toxicity of PEI and increase protection of DNA, the complex of DNA , human serum albumin ( HSA ) and polyethylenimine ( PEI ) was used in this study. pEGFP vector was used as reporter gene to evaluated the transfection of formulations. HSA-PEI-DNA complexes were obtained by adsorbed HSA on PEI-DNA complexes. The complex was also crosslinked by using glutaraldehyde to extend the transfection of DNA. Following two works were conducted in this study, including : (1) To evaluate the physicochemical properties of formulation, such as zeta potential, particle size distribution and surface morphology ; (2) To evaluate the transfect efficiency and toxicity of formulations by using HEK 293 cell model. The jetPEI-DNA complex had the maximal transfection efficiency ( 33.19 % ) which was used as positive control ; the naked pEGFP had the least transfection efficiency ( 0.13 % ) which was used as negative control. In this study we found that HSA could increase the transfect efficiency and decrease the toxicity by adsorbed on the PEI-DNA complex. Three formulations, H5 ( 5 μg DNA ), H 7.5 ( 5 μg DNA ) and H5 ( 10 μg DNA ), showed higher than 20 % cell transfect efficiency and 80 % of cell viability with a good developing value.