装载黄芩苷的溶菌酶-低相对分子质量壳聚糖纳米凝胶的制备及表征

Preparation and characterization of lysozyme-low molecular weight chitosan loaded with baicalin

目的基于蛋白-多糖纳米凝胶的制备技术,通过低相对分子质量壳聚糖对溶菌酶的修饰以及对环境因素的调节筛选,控制溶菌酶的自组装行为,制备一种绿色环保的具备核壳结构的溶菌酶-低相对分子质量壳聚糖纳米凝胶,从而达到减小粒径,提高包封率和载药量的目的,使其更有利于肾靶向。方法利用Maillard反应将溶菌酶与低相对分子质量壳聚糖按不同比例合成溶菌酶-低相对分子质量壳聚糖枝接物,通过SDS-PAGE蛋白电泳来优选出产量最高的合成比例;对合成的产物进行内源荧光、紫外测定等表征,并通过2,4,6-三硝基苯磺酸(2,4,6-trinitro-benzenesulfonic acid,TNBS)法测定接枝率;以粒径为评价指标,对溶菌酶浓度,反应体系pH值、加热温度等条件进行筛选优化,得到空白纳米凝胶最佳合成条件;选取适宜的载药方法,得到了载黄芩苷纳米凝胶。结果合成纳米凝胶的最佳工艺为溶菌酶与低相对分子质量壳聚糖比例1∶2、反应体系pH值11、溶菌酶-壳聚糖接枝物质量浓度为0.6 mg/mL、加热温度71℃、加热时间51 min。得到的溶菌酶-低相对分子质量壳聚糖枝接物接枝率为(24.7±2.9)%,空白纳米凝胶的粒径范围为16~120 nm、平均粒径为49.02 nm、PDI为0.132,载黄芩苷纳米凝胶的包封率为(95.00±2.54)%、载药量为(17.00±1.26)%。结论通过Maillard合成了溶菌酶-低相对分子质量壳聚糖纳米凝胶,并找到了最优的合成工艺。制备的纳米凝胶包封率高,粒径小,分布均匀,缓释效果明显。

Objective Based on the preparation technology of protein polysaccharide nanomaterials,through the modification of low molecular weight chitosan to lysozyme and the adjustment of environmental factors,by controlling the self-assembly behavior of lysozyme,to prepare a green environment-friendly lysozyme low molecular weight chitosan nanomer gel with core shell structure,thereby reducing gel size,increasing entrapment efficiency and drug loading,and making it more conducive to renal target.Methods Lysozyme and low molecular weight chitosan were synthesized by Maillard reaction and SDS-PAGE was used to optimize the ratio of lysozyme and low molecular weight chitosan with the highest yield;The synthesized products were characterized by endogenous fluorescence and UV detection,and the grafting rate was determined by TNBS method;The particle size was used as the evaluation index,The optimal conditions for the preparation of the blank nanomaterials were screened and optimized by optimizing the conditions of lysozyme concentration,reaction system pH and heating temperature.With the entrapment efficiency,the suitable drug loading methods were selected to obtain the nanogel loaded with baicalin.Results The content of lysozyme low molecular weight chitosan with a graft ratio of(24.7±2.9)%was obtained,and the blank nanogel with a particle size of 16—120 nm,an average particle size of 49.02 nm and a PDI 0.132,and a baicalin nanogel with a entrapment efficiency of(95.00±2.54)%and a drug loading of(17.00±1.26)%were obtained.Conclusion Lysozyme low molecular weight chitosan nanoscale gel by Maillard The optimal ratio of lysozyme to low molecular weight chitosan was synthesized by Maillard reaction and the best synthetic process was found.The prepared nano gel has high encapsulation efficiency,small particle size,uniform distribution and obvious sustained release effect.