文献：表面活性蛋白A纳米*体偶联脂质体负载甲泼尼龙提高肺靶向特异性及对急性肺损伤的*效果

链接：https://www.tandfonline.com/doi/full/10.1080/10717544.2017.1402217#abstract

作者：李楠，东翁，王善梅，张元，陈珊珊，尹兆芳，斋嘉丽，朱迪·斯科布尔，夏洛特·C·威廉姆斯，陈涛，邱惠，秦五，赵萌萌，卢立勤，泽维尔·穆莱特&李慧萍

节选：

将 SPANb 与 MPS–NSSLs 共轭形成 MPS–NSSLs–SPANb

将SPANb与MPS-NSSLs混合，然后在4°C下孵育过夜以进行结合。为了优化结合效率，将SPANb和MPS-NSSL以不同的His标签与碘乙酸盐的摩尔比混合。在XCell SureLock Mini-Cell电泳系统（Invitrogen，卡尔斯巴德，加利福尼亚州）上进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）分析，以确认SPANb与MPS-NSSLs的结合。使用Image J软件（Bethesda，MD）对染色的凝胶进行积分，计算结合效率百分比（Gajic等人，引文2011）。结合后，MPS–NSSLs–SPANb（SPANb 和 MPS-NSSLs 的混合物）通过凝胶过滤纯化，纯化柱为 Superose 12 10/300 柱（GE Healthcare，Chicago，IL），并用 PBS 平衡以去除游离的 SPANb。对于 FITC 标记的 MPS-NSSLs-SPANb（MPS-NSSLs-SPANb-FITC），使用相同的方法，将 SPANb 替换为 FITC 标记的 SPANb（SPANb-FITC）。使用 DSPE-PEG-NBD 替换/混合 MPS，然后使用相同方法制备 MPS-NSSLs-NBD/NSSLs-NBD。

Conjugating SPANb to MPS–NSSLs to form MPS–NSSLs–SPANb

SPANb was mixed with MPS–NSSLs and then incubated at 4 °C overnight for conjugation. SPANb and MPS–NSSL were mixed in different mole ratios of His-tag to iodoacetate to optimize conjugation efficiency. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis on an XCell SureLock Mini-Cell Electrophoresis System (Invitrogen, Carlsbad, CA) was performed to confirm the conjugation of SPANb to MPS–NSSLs. Conjugation efficiency in percentage was calculated by integrating the stained gels using Image J software (Bethesda, MD) (Gajic et al., Citation2011). After conjugation, MPS–NSSLs–SPANb (the mixture of SPANb and MPS-NSSLs) was purified by gel filtration on a Superose 12 10/300 column (GE Healthcare, Chicago, IL) equilibrated with PBS to remove of the free SPANb. For the preparation of FITC-labeled MPS-NSSLs-SPANb (MPS-NSSLs-SPANb-FITC), the same protocol was used by replacing SPANb with FITC labeled SPANb (SPANb–FITC). DSPE-PEG-NBD was used to replace/mixed with MPS, and then MPS-NSSLs-NBD/NSSLs-NBD was prepared using the same method.

西安齐岳生物提供相关产品：

GE11-PEG-DSPE

PTP-PEG-DSPE

SP94-PEG-DSPE

T7(HAIYPRH)-PEG-DSPE

Octreotide-PEG-DSPE

YIGSR-PEG-DSPE

APRPG-PEG-DSPE

NGR-PEG-DSPE

NGR(c(GGCNGRC))-PEG-DSPE

RVG29-PEG-DSPE

THRPPMWSPVWP-PEG-DSPE

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