Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory, Livermore, California, United States of America.
PLoS One. 2010 Jul 19;5(7):e11643. doi: 10.1371/journal.pone.0011643.
Nanolipoprotein particles (NLPs) are discoidal, nanometer-sized particles comprised of self-assembled phospholipid membranes and apolipoproteins. NLPs assembled with human apolipoproteins have been used for myriad biotechnology applications, including membrane protein solubilization, drug delivery, and diagnostic imaging. To expand the repertoire of lipoproteins for these applications, insect apolipophorin-III (apoLp-III) was evaluated for the ability to form discretely-sized, homogeneous, and stable NLPs.
Four NLP populations distinct with regards to particle diameters (ranging in size from 10 nm to >25 nm) and lipid-to-apoLp-III ratios were readily isolated to high purity by size exclusion chromatography. Remodeling of the purified NLP species over time at 4 degrees C was monitored by native gel electrophoresis, size exclusion chromatography, and atomic force microscopy. Purified 20 nm NLPs displayed no remodeling and remained stable for over 1 year. Purified NLPs with 10 nm and 15 nm diameters ultimately remodeled into 20 nm NLPs over a period of months. Intra-particle chemical cross-linking of apoLp-III stabilized NLPs of all sizes.
ApoLp-III-based NLPs can be readily prepared, purified, characterized, and stabilized, suggesting their utility for biotechnological applications.
纳米脂蛋白颗粒（NLPs）是圆盘状的纳米级颗粒，由自组装的磷脂膜和载脂蛋白组成。用人类载脂蛋白组装的 NLPs 已被用于无数生物技术应用，包括膜蛋白溶解、药物传递和诊断成像。为了扩大这些应用的脂蛋白库，评估了昆虫载脂蛋白-III（apoLp-III）形成离散大小、均匀和稳定 NLPs 的能力。
通过尺寸排阻色谱法可轻易地从 4 个不同的 NLP 群体中分离出具有不同粒径（范围从 10nm 到 >25nm）和不同脂质与 apoLp-III 比例的高度纯的 NLP 。在 4°C 下，通过天然凝胶电泳、尺寸排阻色谱和原子力显微镜监测纯化的 NLP 物种随时间的重塑情况。纯化的 20nmNLP 没有发生重塑，在超过 1 年的时间里保持稳定。直径为 10nm 和 15nm 的纯化 NLPs 最终在数月内重塑为 20nmNLPs。apoLp-III 内颗粒化学交联稳定了所有大小的 NLPs。
基于 apoLp-III 的 NLPs 可以轻易地制备、纯化、表征和稳定，这表明它们在生物技术应用中的实用性。
