DSPE-PEG-DBCO介导的脂质体表面功能化与生物正构体合成

链接：https://www.researchgate.net/profile/Xiangdong-Xue-2/publication/390580878_Blocked_bioorthogonal_chemistry_enabled_switchable_bioorthosome_to_improve_liposomal_drug_delivery_for_glioblastoma_therapy/links/67f5bfb8e8041142a16fb0dc/Blocked-bioorthogonal-chemistry-enabled-switchable-bioorthosome-to-improve-liposomal-drug-delivery-for-glioblastoma-therapy.pdf

作者：Chao Wang, Jie Wu, Zhiran Duan, Yuqing Pan, Haijing Qu, Wei Cheng, Ning Wang, Han Chen, Xiaoli Gao,Mengqing Hou, Ying Zhang, and Xiangdong Xue

节选：

生物正构体的制备

采用薄膜水合技术制备了生物正构体。具体来说，15毫克大豆的混合物磷脂酰胆碱（PC）、5毫克胆固醇、0.6毫克DSPE-PEG2000，0.3mg DSPE-PEG-PBA和0.1mg DSPE-PEG-DBCO的混合物是溶解在甲醇/氯仿溶液（1:3，v/v）中圆底烧瓶。使用旋转式蒸发器蒸发溶剂蒸发器在烧瓶内表面形成薄脂质膜，随后进行真空干燥，以确保完全去除残留溶剂。所得脂质膜用4mL水复水在超声波处理下，将含有2mg TMZ的PBS溶解。这个然后将再水合的悬浮液通过11次使用Avanti Mini挤出机的聚碳酸酯膜（200 nm）以获得均匀的脂质体。此后，MA（20毫克，0.06毫摩尔）将其加入脂质体溶液中，随后在摇床上孵育过夜以促进化学反应这导致pH响应性硼酸酯键（PBA-MA）的形成，最终导致最终生物正构体的合成。对照脂质体采用相同的方法进行合成，具体来说包括不含pH值的DBCO-MA脂质反应性和生物正交化学屏蔽，以及DBCO-PBA脂质体，其上缺乏葡萄糖功能表面。在体外研究中包封罗丹明B（RhB），而吲哚青绿（ICG）则用于体内所有脂质体组的生物分布分析。

译文：

Preparation of the Bioorthosome

The Bioorthosome was prepared using the thin-film hydrationtechnique. Specifically, a mixture of 15 mg soyphosphatidylcholines (PC), 5 mg cholesterol, 0.6 mg DSPE-PEG2000,0.3 mg DSPE-PEG-PBA, and 0.1 mg DSPE-PEG-DBCO wasdissolved in a methanol/chloroform solution (1:3, v/v) within around-bottom flask. The solvent was evaporated using a rotaryevaporator to form a thin lipid film on the flask's inner surface,followed by vacuum desiccation to ensure complete removal ofresidual solvents. The resulting lipid film was rehydrated with 4 mLof PBS containing 2 mg TMZ under ultrasonication. Therehydrated suspension was then passed 11 times through apolycarbonate membrane (200 nm) using an Avanti Mini-Extruderto obtain uniform liposomes. Thereafter, MA (20 mg, 0.06 mmol)was added to the liposome solution, which was subsequentlyincubated on a shaker overnight to promote the chemical reactionbetween PBA and MA. This resulted in the formation of pHresponsive borate ester bonds (PBA-MA), culminating in thesynthesis of the final Bioorthosome. Control liposomes weresynthesized employing the same methodology, specificallyincluding the DBCO-MA Lipo, which is devoid of pHresponsiveness and bioorthogonal chemistry shielding, as well asthe DBCO-PBA Lipo, which lacks glucose functionality on itssurface. For in vitro studies, rhodamine B (RhB) was encapsulated,while indocyanine green (ICG) was incorporated for in vivobiodistribution analysis across all liposome groups.

西安齐岳生物提供相关产品：

DSPE-PEG-acid

DSPE-PEG4-acid

DSPE-PEG-Ald

DSPE-PEG-Amine，1069-79-0

DSPE-PEG-azide

DSPE-PEG5-azide

DSPE-PEG-Biotin

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