DOPE-PEG-Mal的合成与纯化方法
文献资料：同样传导 CD47 和 PD-L1 可提升遗传性和适合性癌症复发免疫抗体反应迟钝或者生殖细胞分子增加网页链接：//www.thelancet.com/article/S2352-3964(19)30158-6/fulltext作著：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，李嘉节选：MAL-PEG-DOPE的结合MAL-PEG-DOPE的整合基准很多年提出的文章内容。通过EDC/NHS新技术将MAL-PEG-COOH的羧基与DOPE的胺基整合。具体情况方法步骤有以下：将30mg羧基表达的PEG不溶二氯二氧化氮中，并与5mgEDC和4mgNHS混后，室内温度下维持搅伴2h。第二进入8mg DOPE（MAL-PEG-COOH∶DOPE=1∶1，摩尔比），氢气自我保护下现象留宿。将现象代谢物在翻转视频减压蒸馏仪中变干至大方面二氯二氧化氮，第二进入冷乙腈中。未现象的DOPE在2414g下离心式10min，不不溶冷乙腈。上清液在翻转视频减压蒸馏仪中变干为稀脂质。将膜用DD水好多化。将现象代谢物装到透析袋（原子核量= 8 k Da）中，并改变至50 mL DD水溶液中，室内温度下现象48小，离心分离氧化钙含量的EDC/NHS/MAL-PEG-COOH。然后代谢物DOPE-PEG-MAL然后用冻干机冷藏。考虑到核验DOPE-PEG-MAL的整合，对试样确定了核磁振动波谱定量分析。Synthesis of MAL-PEG-DOPESynthesis 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.
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