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文獻名:

Effect of surface charge and density of istearylphosphatidylethanolamine-mPEG-2000 (DSPE-mPEG-2000) on the cytotoxicity of liposome-entrapped ricin: Effect of lysosomotropic agents

文獻鏈接：

https://doi.org/10.1016/j.ijpharm.2007.08.032

實驗使用方法:

脂質體制備

Liposomes composed of soya phosphatidyl choline andcholesterol in a molar ratio of 55:45 were prepared by handshaken method. Briefly, the lipids (40 pmol total lipids)weredissolved in chloroform in a 100ml round bottom fask. Thechloroform was evaporated to dryness at 37°C, under reducedpressure by using rotary evaporator (Wheaton). The thin filmso formed, was desiccated for 1 h, followed by hydration with1 ml PBS (20 mM, pH 7.4), containing ricin (3 mg/ml) and traceamounts of 125]-ricin as aqueous phase marker. The round bot-tom fask containing liposomes suspension was stored, underN2 atmosphere to avoid lipid oxidation, at 4C for overnight forcomplete hydration. The following day, liposomes were soni-cated in a bath type sonicator (Branson) at 25 °C for 30 min in10 min batches to avoid the heat generation. Negatively and positivelycharged liposomes containing ricin were preparedexactlyas described above only 10 mol% either phosphatidic acid (PA)or stearylamine (SA) were added during the preparation of lipidflm. Sterically stabilized liposomes containing ricin were pre-pared as described above by adding various (1-7.5 mol%) ofDSPE-mPEG-2000 during the preparation of lipid film.

重要檢測:

重要實驗結果說明

examined. As shown in Fig. 5 when cells were treated with150 ng/ml free ricin a lag period of 45 min was observed withtso (time required to achieve 50% reduction in protein synthe-sis) of 200 min. It was observed that the lag period of inhibitionof protein synthesis by ricin is significantly increased followingdelivery through different charged liposomes. At 10 pg/ml ofvarious charged liposomal ricin, a lag phase of 4 h was observedfor neutral and negatively charged liposomes, on the other hand.a lag phase of 2h was observed for positively charged lipo-somes. Monensin (50 nm) reduced the lag period of free ricinfrom 45 to 15 min (i.e., three-fold reduction of the lag period),however, in the presence of monensin, the lag period of neutraland negatively chargedliposomal ricin was reduced from 4 to 1 h(four-fold reduction of lag period) and 2 h (two-fold reductionoflag period), respectively. The lag period of positively chargedliposomal ricin was reduced from 2 to 1 h (two-fold reductionof lag period). These results implied that monensin causes anenhanced and efficient release of ricin A-chain from liposomalricin located in an intracellular compartment into the cytosolleading to the rapid onset of the inhibition of protein synthesis.

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關于我們 ：
西安瑞禧生物科技有限公司專業(yè)從事生物醫(yī)藥材料的研發(fā)與生產，提供多種規(guī)格和類型的聚乙二醇（PEG）及其衍生物。其產品涵蓋線性PEG、支鏈PEG、多官能PEG、活性基團修飾PEG（如-COOH、-NH2、-SH、NHS、Mal等）以及脂質修飾類PEG（如DSPE-PEG、Cholesterol-PEG）等，廣泛應用于藥物遞送系統(tǒng)、抗體偶聯(lián)、納米材料修飾、生物成像和組織工程等領域。瑞禧生物可根據(jù)客戶需求定制不同分子量、不同官能團的PEG產品，確保產品質量穩(wěn)定、純度高、反應活性強，滿足科研及產業(yè)化應用需求。


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