文献：Simultaneous blocking of CD47 and PD-L1 increases innate and adaptive cancer immune responses and cytokine release

作者：Shu Liana ∙ Ruizhi Xiea ∙ Yuying Yeb ∙ Xiaodong Xiea ∙ Shuhui Lia ∙ Yusheng Lua,c ∙ et al. Show more

文献链接:https://www.thelancet.com/article/S2352-3964(19)30158-6/fulltext

摘要：

Synthesis of MAL-PEG-DOPE refers to previously published articles. 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.

MAL-PEG-DOPE的合成

MAL-PEG-DOPE的合成参考了之前发表的文章。使用EDC/NHS技术完成了MAL-PEG-COOH羧基与DOPE胺基的偶联。该过程按如下方式进行：30 将mg羧基修饰的PEG溶解在二氯甲烷中，并与EDC（5mg）和NHS（4mg）混合。

将溶液连续搅拌2 h在室温下。随后，8 加入mg DOPE（MAL-PEG-COH:DOPE=1:1，摩尔比），在氮气下反应过夜。反应产物在旋转蒸发器中干燥出大部分二氯甲烷，然后加入冷乙腈中。

将未反应的DOPE在2414g下离心10分钟 min，其不溶于冷乙腈。上清液在旋转蒸发器中干燥成薄脂质。用DD水使薄膜水合。将反应产物装入透析袋（MW = 8 k Da）并转入50 mL DD水溶液，在室温下分离游离的EDC/NNHS/MAL-PEG-COOH 48小时 h.随后用冻干机冷冻最终产物DOPE-PEG-MAL。为了确认DOPE-PEG-MAL共轭，通过核磁共振光谱对样品进行了检测。

相关推荐：

DOPE-PEG-Mal

DOPE-PEG-NH2

DOPE-PEG-SC

DOPE-PEI

DPPE-Cap-Biotin

DPPE-Cap-Folate

DPPE-ICG

DPPE-mPEG

DPPE-PEG2-NH-Mal

以上文章内容来源各类期刊或文献，如有侵权请联系我们删除！