Water-based microsphere delivery system for proteins.
Water-based microsphere delivery system for proteins.
复制标题
用于蛋白质的水基微球输送系统。
DOI：
--
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
T. Speaker
中科院分区：
文献类型：
--
作者：
R. Patil;T. Speaker
本文描述了一种模型蛋白质--辣根过氧化物酶(HRP)在水基微胶囊给药系统中的配方，并展示了这种蛋白质给药系统的实用性。将该酶的水溶液(1 mg/mL)分别与阴离子聚合物IOTA卡拉胶的中性钠盐水溶液(重复单位为0.6 mm)混合。将这些混合物作为均匀的微滴注入到一系列单胺、双胺或寡胺的水溶液中(作为中性盐酸盐或醋酸盐)。基本上，阴离子聚合物的钠盐与胺盐酸盐的瞬时盐交换作用导致了胺/聚合物复合体的微粒的形成。这种酶被捕获在生成的胶囊中。通过反复离心法洗涤，再悬浮于水中，测定其粒度分布。用SDS-PAGE和尺寸排除层析分析洗涤后的微球中HRP的碎裂/聚集，用荧光光谱分析HRP的解折叠，用吸收光谱分析比酶活性、包封率和释放度。根据所用胺的不同，捕获效率从1%到72%不等。胶囊化对蛋白质大小没有不利影响，因为在44 kDa以下或以上看不到分子片段或聚集体。蛋白质的色氨酸荧光光谱在包埋后没有变化，说明三级结构没有构象变化。与底物扩散相关的HRP活性明显降低，但溶解后活性几乎恢复到100%。结果表明，水基荷电薄膜作为蛋白质的药物传递系统，不改变被测蛋白质的结构构象或比活性，并提供恒定速率的蛋白质释放。
This paper describes formulation of a model protein, horseradish peroxidase (HRP), in a water based microcapsule delivery system and demonstrates the utility of this delivery system for proteins. Aqueous solutions (1 mg/mL) of the enzyme were separately blended with aqueous solutions of the neutral sodium salt of the anionic polymer iota carrageenan (0.6 mM in repeat unit). These blends were instilled as uniform microdroplets into aqueous solutions of a series of eleven mono-, di-, or oligo-amines (as neutral hydrochloride or acetate salts). Essentially instantaneous salt exchange interaction of the sodium salt of anionic polymer with amine hydrochloride resulted in formation of microparticles of amine/polymer complex. The enzyme was captured in the resulting capsules. The particles were washed by repeated centrifugation and resuspension in water and their particle size distribution was determined. HRP in washed pelleted microspheres was analyzed for fragmentation/aggregation by SDS-PAGE and size exclusion chromatography, for unfolding by fluorescence spectroscopy, and for specific enzymatic activity, capture efficiency and release studies by absorbance spectroscopy. Dependent on amine employed, capture efficiencies ranged from 1 to 72%. Encapsulation produced no adverse effect on protein size as no molecular fragments or aggregates were visible below or above 44 kDa. The tryptophan fluorescence spectrum of the protein did not change after encapsulation indicating no conformational change in tertiary structure. There was an apparent substrate diffusion related reduction in activity of encapsulated HRP, but almost 100% of activity was recovered on lysis of the capsules. It is concluded that water based charged film encapsulation used as a drug delivery system for proteins does not alter structural conformation or specific activity of the model protein tested and provides protein release at a constant rate.
影响因子：
2.9
作者：
ARAKAWA, T;TIMASHEFF, SN
通讯作者：
TIMASHEFF, SN
影响因子：
5.5
作者：
Gupta, RK;Varanelli, CL;Siber, GR
通讯作者：
Siber, GR
通过微胶囊化增强轮状病毒的免疫原性。
影响因子：
3.7
作者：
Offit,PA;Khoury,CA;Moser,CA;Clark,HF;Kim,JE;Speaker,TJ
通讯作者：
Speaker,TJ