Reversible protein precipitation to ensure stability during encapsulation within PLGA microspheres
Reversible protein precipitation to ensure stability during encapsulation within PLGA microspheres
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DOI：
10.1016/j.ejpb.2008.03.006
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发表时间：
2008-09-01
影响因子：
4.9
通讯作者：
Benoit, Jean-Pierre
中科院分区：
文献类型：
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作者：
Giteau, Alexandra;Venier-Julienne, Marie-Claire;Benoit, Jean-Pierre
使蛋白质沉淀以确保其在随后包封在PLGA微球内时的稳定性。通过向蛋白质溶液中加入盐（氯化钠）和水混溶性有机溶剂（四氢呋喃聚乙二醇醚）形成球形纳米级蛋白质颗粒。通过改变各种工艺参数，优化了溶菌酶、α-糜蛋白酶、过氧化物酶和β-半乳糖苷酶四种模型蛋白的沉淀效率。如通过再水化颗粒的酶活性测量所监测的，对于每种蛋白质定义了获得超过95%的可逆沉淀物的条件。通过吸收光谱、荧光光谱和圆二色性对再水化颗粒的结构的研究表明，沉淀后没有结构扰动。然后使用s/o/w技术将蛋白质颗粒微囊化在PLGA微球内。平均包封产率为约80%，并且在包封步骤后没有发生蛋白质活性的损失。此外，溶菌酶体外释放研究表明，所有释放的溶菌酶都具有生物活性。这种蛋白质沉淀方法适用于各种蛋白质在PLGA微球中的包封而不失活。(c)2008 Elsevier B. V.保留所有权利。
Proteins were precipitated to ensure their stability upon subsequent encapsulation within PLGA microspheres. Spherical, nanosized protein particles were formed by the addition of a salt (sodium chloride) and a water-miscible organic solvent (glycofurol) to protein solutions. Various process parameters were modified to optimize the precipitation efficiency of four model proteins: lysozyme, alpha-chymotrypsin, peroxidase and beta-galactosidase. As monitored by enzymatic activity measurement of the rehydrated particles, conditions to obtain more than 95% of reversible precipitates were defined for each protein. The study of the structure of the rehydrated particles by absorbance spectroscopy, fluorescence spectroscopy and circular dichroism showed an absence of structural-perturbation after precipitation. Protein particles were then microencapsulated within PLGA microspheres using s/o/w technique. The average encapsulation yield was around 80% and no loss of protein activity occurred after the encapsulation step. Additionally, a lysozyme in vitro release study showed that all of the released lysozyme was biologically active. This method of protein precipitation is appropriate for the encapsulation in PLGA microspheres of various proteins without inactivation. (c) 2008 Elsevier B.V. All rights reserved.