文獻(xiàn)：具有增強(qiáng)粘液滲透性的牛奶外泌體可用于口服遞送siRNA

鏈接：https://pubs.rsc.org/en/content/articlehtml/2020/bm/d0bm01497d

作者：Matthew R. Warren a、 Chenzhen 張 a、 Armin Vedadghavami a、 Krister Bokvist b、 Pradeep K. Dhal c和 Ambika G. Bajpayee * ad

節(jié)選：

熒光共軛 DSPE-PEG-疊氮化物的制備。 使用DSPE-PEG-疊氮化物（以下簡(jiǎn)稱DPA）在 mExo 表面形成疏水插入 PEG 涂層。首先將 DPA 以 2 mg mL ?1的濃度溶解在 DMSO 中，然后逐滴添加到 1× PBS 中并不斷攪拌，使 DPA 的最終濃度為 0.02 mg mL ?1（7 μM），從而制備水性儲(chǔ)備溶液。對(duì)于涉及 DPA 濃度量化的研究（即測(cè)量 mExo 表面負(fù)載和錨穩(wěn)定性研究），DPA通過(guò)疊氮化物-DBCO 點(diǎn)擊化學(xué)反應(yīng)與熒光部分 DBCO-Cy5 結(jié)合。將 DBCO-Cy5（DMSO 中濃度為 1 mg mL ?1）逐滴添加到水性 DPA 儲(chǔ)備液中，不斷攪拌，使最終摩爾比為 1 [細(xì)空格（1/6-em）]：[細(xì)空格（1/6-em）]4。然后在室溫下輕輕搖晃該溶液以促進(jìn)點(diǎn)擊反應(yīng)點(diǎn)擊反應(yīng)后，將含有DPA-Cy5和過(guò)量DBCO-Cy5的溶液直接添加到外泌體樣品中進(jìn)行插入。

譯文：

Preparation of fluorescent-conjugated DSPE-PEG-Azide. DSPE-PEG-Azide (hereby DPA) was utilized to enable a hydrophobically-inserted PEG coating on the mExo surface. An aqueous stock solution was prepared by first dissolving DPA in DMSO at 2 mg mL?1, followed by dropwise addition to 1× PBS with continuous stirring for a final DPA concentration of 0.02 mg mL?1 (7 μM). For studies involving quantification of DPA concentration (i.e. measurement of mExo surface loading and anchor stability studies), DPA was conjugated to the fluorescent moiety DBCO-Cy5 via the azide-DBCO click chemistry reaction. DBCO-Cy5 (1 mg mL?1 in DMSO) was added dropwise with continuous stirring to aqueous DPA stock for a 1[thin space (1/6-em)]:[thin space (1/6-em)]4 final molar ratio. This solution was then incubated for 6 h at room temperature with light shaking to facilitate the click reaction. After the click reaction, the solution containing DPA-Cy5 and excess DBCO-Cy5 was added directly to exosome samples for insertion.

西安齊岳生物提供相關(guān)產(chǎn)品：

DSPE-PEG-Cy5

DSPE-PEG-mAb 2C5（二硬脂酰基磷脂酰乙醇胺-聚乙二醇-*靶向蛋白)

DSPE-PEG-OPSS

Cy5-DSPE

DSPE-PEG-Tetrazine

DSPE-PEG-DA

Asp6-PEG-DSPE

DSPE-PEG2000-NGR

Xpeptide-PEG-DSPE

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