文献:同时阻断 CD47 和 PD-L1 可增强先天性和适应性癌症免疫反应以及细胞因子释放
链接:https://www.thelancet.com/article/S2352-3964(19)30158-6/fulltext
作者:舒 莲,谢锐 志,叶玉英,谢晓东,李淑慧,路玉生,李碧飞,程云龙,弗拉基米尔·卡塔纳耶夫,李嘉
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MAL-PEG-DOPE的合成
MAL-PEG-DOPE的合成参考以前发表的文章。采用EDC/NHS技术将MAL-PEG-COOH的羧基与DOPE的胺基结合。具体方法如下:将30mg羧基修饰的PEG溶于二氯甲烷中,并与5mgEDC和4mgNHS混合,室温下连续搅拌2h。然后加入8mg DOPE(MAL-PEG-COOH∶DOPE=1∶1,摩尔比),氮气保护下反应过夜。将反应产物在旋转蒸发仪中干燥至大部分二氯甲烷,然后加入冷乙腈中。未反应的DOPE在2414g下离心10min,不溶于冷乙腈。上清液在旋转蒸发仪中干燥为稀脂质。将膜用DD水水化。将反应产物装入透析袋(分子量= 8 k Da)中,并转移至50 mL DD水溶液中,室温下反应48小时,分离游离的EDC/NHS/MAL-PEG-COOH。最终产物DOPE-PEG-MAL随后用冻干机冷冻。为了验证DOPE-PEG-MAL的结合,对样品进行了核磁共振波谱分析。
Synthesis of MAL-PEG-DOPE
Synthesis of MAL-PEG-DOPE refers to previously published articles [22,23]. The conjugation of carboxyl groups of MAL-PEG-COOH to the amine groups of DOPE was accomplished using the EDC/NHS technique. The process was carried out as follows: 30 mg carboxyl-modified PEG was dissolved in dichloromethane and mixed with EDC (5 mg) and NHS (4 mg). The solution was stirred continuously for 2 h at room temperature. Subsequently, 8 mg DOPE (MAL-PEG-COOH: DOPE =1:1, molar ratio) was added, and the reaction proceeded overnight under nitrogen. The reaction product was dried out most dichloromethane in rotary evaporator and then added to cold acetonitrile. The unreacted DOPE was centrifuged at 2414g for 10 min which was insoluble in cold acetonitrile. And the supernatant was dried to thin lipid in rotary evaporator. The film was hydrated with DD water. The reaction product was enclosed in dialysis bag (MW = 8 k Da) and transferred into 50 mL of DD water solution to separate free EDC/ NHS/ MAL-PEG-COOH at room temperature for 48 h. The final product DOPE-PEG-MAL was subsequently freezed by lyophilizer. To confirm the DOPE-PEG-MAL conjugation, the samples were examined by nuclear magnetic resonance spectroscopy.
西安齐岳生物提供相关产品:
DSPE-PEG-NBD
DSPE-PEG-PDP
DSPE-PEG-Anti-rat CC531mAb(二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)
DSPE-OPSS
DSPE-PEG-SC
LyP-1-PEG-DSPE
Cy7-DSPE
TAT-PEG-DSPE
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