Zhuang F F, Liang R, Zou C T, Ma H, Zheng C X, Duan M X
Department of Biological Science and Biotechnology, Tsinghua University, 100084 Beijing, PR China.
J Biochem Biophys Methods. 2002 Aug-Sep;52(3):169-78. doi: 10.1016/s0165-022x(02)00073-8.
To overcome the drawbacks of encapsulating plasmid DNA (pDNA) in poly (D,L-lactic-co-glycolic acid) (PLGA) by water-in-oil-in-water double-emulsion solvent-evaporation method, we have developed a novel procedure for encapsulating pDNA in PLGA microparticles called DNA organic phase self-emulsification (DOPSM). This method was based on both the extraction plasmid DNA from aqueous phase into organic phase and the spontaneous emulsification DNA in organic phase by solvent diffusion method. The efficiency of extraction plasmid DNA into organic phase is 99% and the concentration of pDNA in organic phase is up to 2.4 mg/ml. The efficiency of microencapsulation of plasmid DNA in PLGA is up to 76% and can be enhanced by lowering the pH of aqueous solution of emulsion. The microparticles size of PLGA of pDNA is in a narrow range of 1-2 microm. This procedure does not involve the high mechanical energy to emulsify which may damage the integrity of pDNA. This method can be applied to encapsulate the pDNA into microparticles of other biocompatible polymers with high efficiency.
为克服采用水包油包水双乳液溶剂蒸发法将质粒DNA（pDNA）包裹于聚（D，L-乳酸-乙醇酸）（PLGA）中存在的缺点，我们开发了一种将pDNA包裹于PLGA微粒中的新方法，称为DNA有机相自乳化法（DOPSM）。该方法基于从水相将质粒DNA萃取至有机相以及通过溶剂扩散法使DNA在有机相中自发乳化。将质粒DNA萃取至有机相的效率为99%，有机相中pDNA的浓度高达2.4 mg/ml。pDNA在PLGA中的微囊化效率高达76%，且可通过降低乳液水溶液的pH值来提高。pDNA的PLGA微粒尺寸在1 - 2微米的窄范围内。该方法不涉及可能破坏pDNA完整性的高机械能乳化过程。此方法可高效应用于将pDNA包裹于其他生物相容性聚合物的微粒中。
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