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生物PISA中囊泡性质的表征与优化：从尺寸分布到组装后加载

Characterization and Optimization of Vesicle Properties in bioPISA: from Size Distribution to Post-Assembly Loading.

作者信息

Belluati Andrea, Bloch Adrian, Koynov Kaloian, Müller Nieva Mariana, Bagherabadi Mohadeseh, Andrieu-Brunsen Annette, Kolmar Harald, Bruns Nico

机构信息

Department of Pure and Applied Chemistry, University of Strathclyde, Thomas Graham Building, 295 Cathedral Street, Glasgo, G1 1XL, UK.

Department of Chemistry, Technical University of Darmstadt, Peter-Grünberg-Straße 4, 64287, Darmstadt, Germany.

出版信息

Adv Biol (Weinh). 2025 May;9(5):e2400483. doi: 10.1002/adbi.202400483. Epub 2024 Dec 18.

DOI:10.1002/adbi.202400483
PMID:39692631
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12078863/
Abstract

This study investigates the formation and properties of vesicles produced via biocatalytic Polymerization-Induced Self-Assembly (bioPISA) as artificial cells. Methods for achieving size uniformity, including gentle centrifugation and sucrose gradient centrifugation, are explored, and the effects of stirring speed on vesicle morphology is investigated. The internal structure of the vesicles, characterized by a polymer-rich matrix, is analyzed using fluorescence correlation spectroscopy (FCS). Additionally, the feasibility of loading macromolecules into pre-formed vesicles is demonstrated using electroporation, and a fluorescent protein as well as enzymes for a cascade reaction were sucesfully incorporated into the fully assembled polymersomes. These findings provide a foundation for developing enzyme-synthesized polymeric vesicles with controlled morphologies for various applications, e.g., in synthetic biology.

摘要

本研究调查了通过生物催化聚合诱导自组装（bioPISA）产生的囊泡作为人工细胞的形成及性质。探索了实现尺寸均匀性的方法，包括温和离心和蔗糖梯度离心，并研究了搅拌速度对囊泡形态的影响。使用荧光相关光谱（FCS）分析了以富含聚合物的基质为特征的囊泡内部结构。此外，通过电穿孔证明了将大分子加载到预先形成的囊泡中的可行性，并且一种荧光蛋白以及用于级联反应的酶成功地掺入了完全组装的聚合物囊泡中。这些发现为开发具有可控形态的酶合成聚合物囊泡用于各种应用（例如合成生物学）奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/16876454adf8/ADBI-9-2400483-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/5b77fff6b9e8/ADBI-9-2400483-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/ece44c905534/ADBI-9-2400483-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/98e749079372/ADBI-9-2400483-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/acfcbe69298d/ADBI-9-2400483-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/04b39fcd9054/ADBI-9-2400483-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/5ba02dc73f95/ADBI-9-2400483-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/b09e41a6b11a/ADBI-9-2400483-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/16876454adf8/ADBI-9-2400483-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/5b77fff6b9e8/ADBI-9-2400483-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/ece44c905534/ADBI-9-2400483-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/98e749079372/ADBI-9-2400483-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/acfcbe69298d/ADBI-9-2400483-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/04b39fcd9054/ADBI-9-2400483-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/5ba02dc73f95/ADBI-9-2400483-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/b09e41a6b11a/ADBI-9-2400483-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a627/12078863/16876454adf8/ADBI-9-2400483-g009.jpg

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本文引用的文献

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Artificial cell synthesis using biocatalytic polymerization-induced self-assembly.
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An Outer Membrane-Inspired Polymer Coating Protects and Endows Escherichia coli with Novel Functionalities.一种受外膜启发的聚合物涂层可保护大肠杆菌并赋予其新的功能。
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