Department of Chemical and Biological Engineering, Iowa State University, Ames, IA 50011, USA.
Acta Biomater. 2010 Aug;6(8):3110-9. doi: 10.1016/j.actbio.2010.01.040. Epub 2010 Feb 1.
The design of biodegradable polymeric delivery systems based on polyanhydrides that would provide for improved structural integrity of Yersinia pestis antigens was the main goal of this study. Accordingly, the full-length Y. pestis fusion protein (F1-V) or a recombinant Y. pestis fusion protein (F1(B2T1)-V10) was encapsulated and released from microparticles based on 1,6-bis(p-carboxyphenoxy)hexane (CPH) and sebacic acid (SA) copolymers and 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) and CPH copolymers fabricated by cryogenic atomization. An enzyme-linked immunosorbent assay was used to measure changes in the antigenicity of the released proteins. The recombinant F1(B2T1)-V10 was unstable upon release from the hydrophobic CPH:SA microparticles, but maintained its structure and antigenicity in the amphiphilic CPTEG:CPH system. The full-length F1-V was stably released by both CPH:SA and CPTEG:CPH microparticles. In order to determine the effect of the anhydride monomers on the protein structure, changes in the primary, secondary, and tertiary structure, as well as the antigenicity of both Y. pestis antigens, were measured after incubation in the presence of saturated solutions of SA, CPH, and CPTEG anhydride monomers. The results indicated that the amphiphilic environment provided by the CPTEG monomer was important to preserve the structure and antigenicity of both proteins. These studies offer an approach by which a thorough understanding of the mechanisms governing antigenic instability can be elucidated in order to optimize the in vivo performance of biodegradable delivery devices as protein carriers and/or vaccine adjuvants.
本研究的主要目标是设计基于聚酸酐的可生物降解聚合物递药系统，以提高鼠疫耶尔森氏菌抗原的结构完整性。因此，全长鼠疫耶尔森氏菌融合蛋白（F1-V）或重组鼠疫耶尔森氏菌融合蛋白（F1(B2T1)-V10）被包裹在基于 1,6-双（对羧基苯氧基）己烷（CPH）和癸二酸（SA）共聚物以及 1,8-双（对羧基苯氧基）-3,6-二氧杂辛烷（CPTEG）和 CPH 共聚物的微球中，并通过低温雾化来制备。酶联免疫吸附试验用于测量释放蛋白的抗原性变化。重组 F1(B2T1)-V10 在从疏水性 CPH:SA 微球中释放时不稳定，但在亲脂性 CPTEG:CPH 系统中保持其结构和抗原性。全长 F1-V 可稳定地从 CPH:SA 和 CPTEG:CPH 微球中释放。为了确定酸酐单体对蛋白质结构的影响，在饱和 SA、CPH 和 CPTEG 酸酐单体溶液存在下孵育后，测量了两种鼠疫耶尔森氏菌抗原的一级、二级和三级结构以及抗原性的变化。结果表明，CPTEG 单体提供的两亲环境对于保持两种蛋白质的结构和抗原性很重要。这些研究提供了一种方法，可以深入了解控制抗原不稳定性的机制，从而优化可生物降解递药系统作为蛋白质载体和/或疫苗佐剂的体内性能。
