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A promising approach toward efficient isolation of the exosomes by core-shell PCL-gelatin electrospun nanofibers.
Bioprocess and Biosystems Engineering ( IF 3.5 ) Pub Date : 2020-07-01 , DOI: 10.1007/s00449-020-02385-7
Fatemeh Barati 1 , Arezoo Mohammadian Farsani 1 , Matin Mahmoudifard 1
Affiliation  

Exosomes as cell-derived vesicles are promising biomarkers for noninvasive and early detection of different types of cancer. However, a straightforward and cost-effective technique for isolation of exosomes in routine clinical settings is still challenging. Herein, we present for the first time, a novel coaxial nanofiber structure for the exosome isolation from body fluids with high efficiency. Coaxial nanofiber structure is composed of polycaprolactone polymer as core and a thin layer of gelatin (below 10 nm) as the shell. The thermo-sensitive thin layer of gelatin can efficiently release the captured exosome by specific antibody namely, CD63, whenever its temperature raised to the physiological temperature of 37 °C. Moreover, the thin layer of gelatin induces less contamination to separated exosomes. The interconnected micro-pores of electrospun nanofibrous membrane insurances large surface area for immobilization of specific antibody for efficient exosome capturing. The efficacy of exosome isolation is determined by direct ELISA and compared with ultracentrifugation technique. For the exosome isolation, it was observed that over 87% of exosomes existed in the culture medium can be effectively isolated by coaxial electrospun nanofibers with the average thickness of 50 µm. Therefore, this promising technique can be substituted for the traditional techniques for exosome isolation which are mostly suffering from low efficacy, high cost, and troublesome process.



中文翻译:

通过核-壳 PCL-明胶电纺纳米纤维有效分离外泌体的一种有前途的方法。

作为细胞衍生囊泡的外泌体是用于无创和早期检测不同类型癌症的有前途的生物标志物。然而,在常规临床环境中分离外泌体的直接且具有成本效益的技术仍然具有挑战性。在此,我们首次提出了一种新型同轴纳米纤维结构,用于高效地从体液中分离外泌体。同轴纳米纤维结构由聚己内酯聚合物为核,明胶薄层(10nm以下)为壳。热敏性明胶薄层可以通过特异性抗体 CD63 有效释放捕获的外泌体,只要其温度升高到 37°C 的生理温度。此外,明胶薄层对分离的外泌体的污染更少。电纺纳米纤维膜的互连微孔确保了大表面积,用于固定特异性抗体,以实现有效的外泌体捕获。外泌体分离的功效是通过直接 ELISA 确定的,并与超速离心技术进行比较。对于外泌体分离,观察到培养基中存在的 87% 以上的外泌体可以通过平均厚度为 50 µm 的同轴电纺纳米纤维有效分离。因此,这种有前景的技术可以替代传统的外泌体分离技术,这些技术主要是效率低、成本高、过程麻烦。外泌体分离的功效是通过直接 ELISA 确定的,并与超速离心技术进行比较。对于外泌体分离,观察到培养基中存在的 87% 以上的外泌体可以通过平均厚度为 50 µm 的同轴电纺纳米纤维有效分离。因此,这种有前景的技术可以替代传统的外泌体分离技术,这些技术主要是效率低、成本高、过程麻烦。外泌体分离的功效是通过直接 ELISA 确定的,并与超速离心技术进行比较。对于外泌体分离,观察到培养基中存在的 87% 以上的外泌体可以通过平均厚度为 50 µm 的同轴电纺纳米纤维有效分离。因此,这种有前景的技术可以替代传统的外泌体分离技术,这些技术主要是效率低、成本高、过程麻烦。

更新日期:2020-07-01
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