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目的研制白蛋白纳米基因载体,评估白蛋白纳米粒用作基因载体的可行性。方法采用去溶剂化法制备白蛋白纳米粒,并通过控制转速、pH值、搅拌时间三个变量来考察白蛋白纳米粒的表征变化情况,以PicoGreen荧光法来测白蛋白纳米载体的基因转载率,并用流式细胞仪检测膀胱癌细胞的基因转染率。结果控制转速1000RPM,自然状态下的溶液pH值(6.9),持续搅拌12h条件下,制得满足基因载体需要的白蛋白纳米粒的平均粒径(253.1±11.9)nm,zeta电位(-34.0±4.5)mV,PDI0.43±0.04,性质相对稳定,基因转载率为(97.61±0.06)%,转染率为(48.21±5.94)%。结论单纯质粒很难进入细胞核高表达,借助白蛋白纳米载体,使高效的基因转染成为可能。白蛋白纳米粒有望被广泛应用为二次基因转载工具。
OBJECTIVE: To develop an albumin nanoparticle vector and evaluate the feasibility of using albumin nanoparticles as a gene vector. Methods The albumin nanoparticles were prepared by desolvation method. The changes of the characterization of albumin nanoparticles were investigated by controlling the rotating speed, pH value and stirring time. The gene transfer rate of albumin nanocarriers was measured by PicoGreen fluorescence method. , And the gene transfection rate of bladder cancer cells was detected by flow cytometry. Results The average diameter of albumin nanoparticles (253.1 ± 11.9) nm and the zeta potential (-34.0 ± 10) were obtained under the conditions of 1000RPM rotational speed, 6.9% natural pH value and 12 hours continuous stirring. 4.5) mV and PDI0.43 ± 0.04. The gene transfer rate was (97.61 ± 0.06)% and the transfection rate was (48.21 ± 5.94)%. Conclusion Simple plasmid is difficult to enter the high expression of the nucleus, with the help of albumin nanocarriers, so that efficient gene transfection as possible. Albumin nanoparticles are expected to be widely used as secondary gene transfer tools.