Protein stability in the presence of polymer degradation products: Consequences for controlled release formulations
Protein stability in the presence of polymer degradation products: Consequences for controlled release formulations
复制标题
DOI：
10.1016/j.biomaterials.2006.01.054
复制
发表时间：
2006-06-01
期刊：
影响因子：
14
通讯作者：
Narasimhan, B
中科院分区：
文献类型：
--
作者：
Determan, AS;Wilson, JH;Narasimhan, B
当将蛋白质包封在聚合物微球中用于持续药物递送时，存在三个阶段，在这三个阶段期间必须保持蛋白质的稳定性：（1）微球的制造，（2）微球的储存，和（3）包封的蛋白质的释放。本研究的重点是在酯（乳酸和乙醇酸）和酸酐（癸二酸和1.6-双（对羧基苯氧基）己烷）单体存在下孵育0或20天后，聚合物降解产物对破伤风类毒素、卵清蛋白（Ova）和溶菌酶的一级、二级和三级结构的影响。在每种单体存在下，定量每种蛋白质的结构和抗原性或酶活性。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，圆二色性，荧光光谱分别用于评估/评价蛋白质的一级，二级和三级结构。用酶联免疫吸附试验测定破伤风类毒素和卵蛋白的抗原性变化，用荧光法测定溶菌酶的酶活性。破伤风类毒素被认为是最稳定的酸酐单体的存在下，而卵是最稳定的癸二酸的存在下，和溶菌酶是稳定的，当孵育的所有单体的研究。(c)2006爱思唯尔有限公司保留所有权利。
When encapsulating proteins in polymer microspheres for sustained drug delivery there are three stages during which the stability of the protein must be maintained: (1) the fabrication of the microspheres, (2) the storage of the microspheres, and (3) the release of the encapsulated protein. This Study focuses on the effects of polymer degradation products on the primary, secondary, and tertiary structure of tetanus toxoid, ovalbumin (Ova), and lysozyme after incubation for 0 or 20 days in the presence of ester (lactic acid and glycolic acid) and anhydride (sebacic acid and 1.6-bis(p-carboxyphenoxy)hexane) monomers. The structure and antigenicity or enzymatic activity of each protein in the presence of each monomer was quantified. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, circular dichroism, and fluorescence spectroscopy were used to assess/evaluate the primary, secondary, and tertiary structures of the proteins, respectively. Enzyme-linked immunosorbent assay was used to measure changes in the antigenicity of tetanus toxoid and Ova and a fluorescence-based assay Was used to determine the enzymatic activity of lysozyme. Tetanus toxoid was found to be the most stable in the presence of anhydride monomers, while Ova was most stable in the presence of sebacic acid, and lysozyme was stable when incubated with all of the monomers studied. (c) 2006 Elsevier Ltd. All rights reserved.