文獻(xiàn)：Exosomal Vaccine Loading T Cell Epitope Peptides of SARS-CoV-2 Induces Robust CD8+ T Cell Response in HLA-A Transgenic Mice

作者：An-Ran Shen 1, Xiao-Xiao Jin 2, Tao-Tao Tang 1, Yan Ding 2, Xiao-Tao Liu 2, Xin Zhong 1, Yan-Dan Wu 2, Xue-Lian Han 3, Guang-Yu Zhao 3, Chuan-Lai Shen 2, Lin-Li Lv 1,?, Bi-Cheng Liu 1

文獻(xiàn)鏈接：https://pmc.ncbi.nlm.nih.gov/articles/PMC9346304/

摘要：

We used the same procedure used for generating the BSA-PEG-lipid-exosome complex to generate the peptide-PEG-lipid-exosome complex. DMPE-PEG-NHS and the mixture of VEPs were dissolved in DMSO at a 1:3 molar ratio with 1% TEA overnight at RT to form the peptides-PEG-DMPE complex. Then, HEPES buffer was added to dilute the DMSO concentration below 1%. Then, DSPE-PEG was added at a 1:1 molar ratio to the pre-existing peptides-DMPE-PEG-NHS complex and maintained for 15 min at 60℃. The free peptides were removed, and the protein levels in the resultant peptides-PEG-micelles were quantified using the BCA assay kit and stored at 4℃ for use in the following 2 weeks. Then, the peptides-PEG-micelles were redissolved, ultrasonicated to reduce the micelle size, and co-incubated with the RBC-derived exosomes at a 1:1 ratio in a water bath for 2 h at 40℃. The resulting solution was cooled to 4℃ followed by instant purification by the FPLC system with the SXK16/40 Smartarose CL-4B filtration column. Finally, the fractions of the second peak were collected, concentrated in normal saline by ultrafiltration, and used as the peptides-PEG-lipid-exosome complex.

我們使用與產(chǎn)生BSA-PEG脂質(zhì)外泌體復(fù)合物相同的程序來產(chǎn)生肽-PEG-脂質(zhì)外泌物復(fù)合物。

將DMPE-PEG-NHS和VEP的混合物以1:3的摩爾比與1%TEA在室溫下溶解在DMSO中過夜，形成肽-PEG-DMPE復(fù)合物。

然后，加入HEPES緩沖液以將DMSO濃度稀釋至1%以下。然后，將DSPE-PEG以1:1的摩爾比加入到預(yù)先存在的肽DMPE-PEG-NHS復(fù)合物中，并在60℃下保持15分鐘。

去除游離肽，使用BCA檢測(cè)試劑盒定量所得肽-PEG膠束中的蛋白質(zhì)水平，并將其儲(chǔ)存在4℃下，以便在接下來的2周內(nèi)使用。

然后，將肽-PEG膠束重新溶解，超聲處理以減小膠束尺寸，并在40℃的水浴中以1:1的比例與RBC衍生的外泌體共孵育2小時(shí)。

將所得溶液冷卻至4℃，然后用SXK16/40 Smartrose CL-4B過濾柱通過FPLC系統(tǒng)立即純化。最后，收集第二個(gè)峰的組分，通過超濾在生理鹽水中濃縮，用作肽-PEG-脂質(zhì)外泌體復(fù)合物。

相關(guān)推薦：

DSPE-Chitosan

DSPE-RB

DSPE-FITC

DSPE-ICG

DSPE-OPSS

DSPE-PCL

DSPE-PEI

DSPE-PEI-Dextran

DSPE-PLA

DSPE-PLGA

DSPE-Poly(L-glutamic acid)

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