AbstractMembrane proteins tend to be difficult to study since they need to be integrated into a lipid bilayer membrane to function properly. This study presents a method to synthesize a macroscopically large and freely transportable membrane with integrated membrane proteins which is useful for studying membrane proteins and protein complexes in isolation.

摘要膜蛋白往往难以研究，因为它们需要整合到脂质双层膜中才能正常工作。这项研究提出了一种合成具有整合膜蛋白的宏观大且可自由运输的膜的方法，该方法可用于分离研究膜蛋白和蛋白质复合物。

The method could serve as a blueprint for the production of larger quantities of functionalised membranes for integration into technical devices similar to the MinION DNA sequencer. It is possible to self-assemble larger biological membranes on solid surfaces. However, they cannot be removed from their solid support without destroying them.

该方法可以作为生产大量功能化膜的蓝图，以整合到类似于MinION DNA测序仪的技术设备中。可以在固体表面上自组装较大的生物膜。然而，如果不摧毁它们，它们就不能从其坚实的支撑物中移除。

In transportable form, self-assembled membranes are limited to sizes of about 17 nm in nanodiscs. Here we electrospray a series of molecular layers onto the liquid surface of a buffer solution which creates a flat, liquid environment on the surface that directs the self-assembly of the membrane. This method enables us to experimentally control the membrane composition and to succeed in producing large membranes with integrated OmpG, a transmembrane pore protein.

以可运输的形式，自组装膜在纳米盘中的尺寸限制在约17 nm。在这里，我们将一系列分子层电喷雾到缓冲溶液的液体表面上，在表面上产生平坦的液体环境，指导膜的自组装。这种方法使我们能够通过实验控制膜的组成，并成功地生产出具有整合的OmpG（一种跨膜孔蛋白）的大型膜。

The technique is compatible with the assembly of membrane based protein complexes. Listeriolysin O and pneumolysin efficiently assemble into non-covalent membrane pore complexes of approximately 30 units or more within the surface layer..

该技术与基于膜的蛋白质复合物的组装兼容。李斯特菌溶血素O和溶血素在表面层内有效组装成约30个单位或更多的非共价膜孔复合物。。

IntroductionThe aim of this study is to make membranes with integrated proteins accessible for functional and structural studies. We are using a nanoelectrospray device to synthesize an environment from gas phase in which the membranes can self-assemble. The molecular composition of the membrane, its lipid and proteinaceous components, is controlled by the experimental design.We use a nanoelectrospray ion source to create a planar environment for self-assembly of large lipid bilayers containing membrane proteins.

引言本研究的目的是使具有整合蛋白的膜可用于功能和结构研究。我们正在使用纳米电喷雾装置从气相合成环境，在其中膜可以自组装。膜的分子组成，其脂质和蛋白质成分，由实验设计控制。。

In order to maintain the proteins’ three-dimensional shape, the technique was conducted under atmospheric conditions and ambient temperatures (see Fig. 1 panel a)1.Fig. 1Electrospray apparatus for surface preparation. Panel (a) shows the electrospray apparatus for membrane preparation (2nd generation instrument): A pressurized container holds a gold-plated, drawn glass capillary filled with 1 or 2 µL solution (1).

为了保持蛋白质的三维形状，该技术是在大气条件和环境温度下进行的（见图1图a）1。图1用于表面制备的电子射线装置。图（a）显示了用于膜制备的电喷雾装置（第二代仪器）：加压容器装有一个镀金的拉制玻璃毛细管，其中填充有1或2µL溶液（1）。

An optional low pressure supports the continuous flow of sample through the glass capillary during operation. The distance between the grounded target (2) and the needle is adjustable. It was usually 3 cm. The first generation instrument had a 250 μm steel nozzle and a manually drawn glass capillary as the emitter.

可选的低压支持样品在操作过程中连续流过玻璃毛细管。接地目标（2）和针之间的距离是可调的。它通常是3厘米。第一代仪器具有250μm的钢喷嘴和手动绘制的玻璃毛细管作为发射器。

Its vessel was not pressurized7. Panel (b) shows how this instrument was used to create a layered environment for the self-assembly of large membranes with membrane proteins. The spray was directed at the liquid surface in a conductive container filled with buffer solution with some SM2 beads at the bottom.

其容器未加压7。图（b）显示了如何使用该仪器为大膜与膜蛋白的自组装创造分层环境。喷雾被引导到填充有缓冲溶液的导电容器中的液体表面，底部有一些SM2珠。

First, a lipid bilayer was formed by spraying the appropriate amounts of phosphatidylcholine. A thin layer of glycerol was then added. Finally, the detergent-solubilized membrane protein was added with some lipid to form a closed membran.

首先，通过喷洒适量的磷脂酰胆碱形成脂质双层。然后加入一薄层甘油。最后，将去污剂溶解的膜蛋白加入一些脂质以形成封闭的膜。

(1)

(1)

re: emission radius of droplets at the tip of the Taylor cone, ρ: density of the solution, γ: surface tension, ϑ: opening angle of the Taylor cone (49.3° in the static case ), Ua: applied and UT Taylor cone threshold voltage, \(\dot{V}\)flow rate.Fig. 2Electrospray prepared surface.

re：泰勒锥尖端液滴的发射半径，ρ：溶液密度，γ：表面张力，ϑ：泰勒锥的张开角（静态情况下为49.3°），Ua：施加的和UT泰勒锥阈值电压，\（\ dot{V}）流速。图2电子射线处理表面。

Panel (a) shows the raster electron microscopic image of a surface prepared by electrospraying from a 250 μm nozzle. The surface layer was deposited from a 30 µM rhodamine acetone solution sprayed onto a metal target at a flow rate of 8–15 µL/min. The image shows a 10 000x magnification of the gold-coated rhodamine surface.

图（a）显示了通过从250μm喷嘴电喷雾制备的表面的光栅电子显微镜图像。表面层是从以8-15µL/min的流速喷涂到金属靶材上的30µM罗丹明丙酮溶液中沉积而成的。该图像显示了镀金罗丹明表面的10000倍放大倍数。

The surface is densely covered with 0.2–1.5 μm particles. Panel (b) shows that the maximum observed particle size on the target depends on the flow rate in accordance with formula 1. When spraying a saturated rhodamine/acetone solution, the largest particle diameter on the target is approximately the size of the initial droplets.

表面密布着0.2-1.5μm的颗粒。图（b）显示，根据公式1，目标上观察到的最大粒径取决于流速。当喷洒饱和罗丹明/丙酮溶液时，目标上的最大粒径约为初始液滴的大小。

Panel (b) had been published before when describing the electrospray theory7. Panel (c) shows a layer prepared with rhodamine using a self-sustained flow rate of approximately 200 nL/min generated with a hand-pulled gold-coated glass capillary. The photo shows the surface magnified 20 000 times. While rhodamine covered the entire surface (see panel d), no specific particle sizes could be determined.

小组（b）在描述电喷雾理论7之前已经发表过。图（c）显示了用罗丹明制备的层，使用手拉镀金玻璃毛细管产生的约200 nL/min的自持流速。照片显示表面放大了2万倍。虽然罗丹明覆盖了整个表面（见图d），但无法确定特定的粒径。

Panel (d) shows the static SIMS signal of the same target as in panel (c), but before it was coated with gold. The spatially resolved static SIMS showed that the rhodamine uniformly covered a 4 mm wide area. Panel (e) shows how the static SIMS Rh+ signal as a function of the amount of rhodamine sprayed onto the surface.

图（d）显示了与图（c）中相同目标的静态SIMS信号，但在它被镀金之前。空间分辨静态SIMS显示罗丹明均匀覆盖4毫米宽的区域。图（e）显示了静态SIMS Rh+信号如何与喷洒在表面上的罗丹明量有关。

The liquid sprayed was a 30 µM rhodamine-acetone solution. For each target, three different measurements at differe.

喷雾的液体是30µM罗丹明丙酮溶液。对于每个目标，在不同的位置进行三次不同的测量。

Data availability

数据可用性

All relevant electron microscopic images are part of the article or its supplemental information. The overlap of the OmpG structure model derived from its crystal structure with the electron microscopic image has been made available via figshare (https://doi.org/10.6084/m9.figshare.27074134) and the movie is part of the supplemental files..

所有相关的电子显微镜图像都是文章或其补充信息的一部分。通过figshare可以获得由其晶体结构得出的OmpG结构模型与电子显微镜图像的重叠(https://doi.org/10.6084/m9.figshare.27074134)这部电影是补充文件的一部分。。

AbbreviationsOmpG:

缩写OmpG：

outer membrane protein G

外膜蛋白G

E. coli:

E、 大肠杆菌：

Escherichia coli

大肠杆菌

TOF-SIMS:

TOF-SIMS：

Time of flight - secondary Ion mass spectrometry

飞行时间-二次离子质谱法

ReferencesWilm, M. Synthesis of Extended, Self-assembled Biological membranes containing membrane proteins from gas phase. bioRxiv 23, 661215 (2019).

参考文献Wilm，M。合成含有气相膜蛋白的扩展自组装生物膜。bioRxiv 23661215（2019）。

Google Scholar

谷歌学者

Yang, Z. et al. Electrospray-assisted cryo-EM sample preparation to mitigate interfacial effects. Nat. Methods (2024).Giess, F., Friedrich, M. G., Heberle, J., Naumann, R. L. & Knoll, W. The protein-tethered lipid bilayer: a Novel Mimic of the Biological membrane. Biophys. J. 87, 3213–3220 (2004).Article .

Yang，Z.等人。电喷雾辅助低温电磁样品制备以减轻界面效应。自然方法（2024）。Giess，F.，Friedrich，M.G.，Heberle，J.，Naumann，R.L。＆Knoll，W。蛋白质束缚的脂质双层：生物膜的新型模拟物。生物物理。J、 873213-3220（2004）。文章。

ADS

广告

CAS

中科院

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

De Zorzi, R., Nicholson, W. V., Guigner, J. M., Erne-Brand, F. & Vénien-Bryan, C. Growth of large and highly ordered 2D crystals of a K⁺ channel, structural role of lipidic environment. Biophys. J. 105, 398–408 (2013).Article

De Zorzi，R.，Nicholson，W.V.，Guigner，J.M.，Erne Brand，F。＆Vénien Bryan，C。K+通道的大而高度有序的2D晶体的生长，脂质环境的结构作用。生物物理。J、 105398-408（2013）。文章

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Denisov, I. G. & Sligar, S. G. Nanodiscs in membrane biochemistry and biophysics. Chem. Rev. 117, 4669–4713 (2017).Article

Denisov，I.G。＆Sligar，S.G。膜生物化学和生物物理学中的纳米盘。化学。修订版1174669–4713（2017）。文章

CAS

中科院

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Jaworek, A. Electrospray droplet sources for thin film deposition. J. Mater. Sci. 42, 266–297 (2007).Article

Jaworek，A。用于薄膜沉积的电喷雾液滴源。J、 马特。科学。42266-297（2007）。文章

ADS

广告

CAS

中科院

Google Scholar

谷歌学者

Wilm, M. & Mann, M. Electrospray and Taylor-Cone theory, Dole’s beam of macromolecules at last? Int. J. Mass Spectrom. Ion Process. 136, 167–180 (1994).Article

Wilm，M。＆Mann，M。电喷雾和泰勒锥理论，Dole的大分子束终于出现了？。离子过程。136167-180（1994）。文章

ADS

广告

CAS

中科院

Google Scholar

谷歌学者

Wilm, M. Principles of electrospray ionization. Molecular & cellular proteomics: MCP 10, M111.009407 (2011).Benninghoven, A., Hagenhoff, B. & Niehuis, E. Surface MS: probing real-world samples. Anal. Chem. 65, 630A–640A (1993).Article

Wilm，M。电喷雾电离原理。分子与细胞蛋白质组学：MCP 10，M111.009407（2011）。。肛门。化学。65630A–640A（1993）。文章

CAS

中科院

Google Scholar

谷歌学者

Wilm, M. & ter Meer-Müller-Steinmeister and Partner, E. P. A., Artur-Ladebeck-Str. 51, D-33617 Bielefeld. Das Micro-Elektrospray (1991).CKMeng, Mann, M. & Fenn, J. B. Of protons or proteins. Z. für Physik D Atoms Molecules Clusters 10, 361–368 (1988).Article

Wilm，M.&ter Meer-Müller-Steinmeister，合伙人，E.P.A.，Artur-Ladebeck-Str.51，D-33617比勒费尔德。微电喷雾（1991）。CKMeng，Mann，M.和Fenn，J.B.关于质子或蛋白质。Z.为物理学D原子分子团10，361-368（1988）。文章

ADS

广告

Google Scholar

谷歌学者

Fenn, J. B., Mann, M., Meng, C. K., Wong, S. F. & Whitehouse, C. M. Electrospray ionization for mass spectrometry of large biomolecules. Science (New York NY) 246, 64–71 (1989).Article

Fenn，J.B.，Mann，M.，Meng，C.K.，Wong，S.F。和Whitehouse，C.M。电喷雾电离用于大生物分子的质谱分析。《科学》（纽约州纽约）246,64-71（1989）。文章

ADS

广告

CAS

中科院

Google Scholar

谷歌学者

Fenn, J. Ion Formation from Charged Droplets: Roles of Geometry, Energy, and time (Journal of the American Society for Mass Spectrometry, 1993).Mann, M. & Wilm, M. Electrospray mass spectrometry for protein characterization. Trends Biochem. Sci. 20, 219–224 (1995).Article

Fenn，J。带电液滴的离子形成：几何，能量和时间的作用（美国质谱学会杂志，1993）。Mann，M。＆Wilm，M。电喷雾质谱法用于蛋白质表征。趋势生物化学。科学。20219-224（1995）。文章

CAS

中科院

PubMed

PubMed

Google Scholar

谷歌学者

Yildiz, O., Vinothkumar, K. R., Goswami, P. & Kühlbrandt, W. Structure of the monomeric outer-membrane porin OmpG in the open and closed conformation. EMBO J. 25, 3702–3713 (2006).Article

Yildiz，O.，Vinothkumar，K.R.，Goswami，P。＆Kühlbrandt，W。开放和闭合构象中单体外膜孔蛋白OmpG的结构。EMBO J.253702–3713（2006）。文章

CAS

中科院

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Subbarao, G. V. & van den Berg, B. Crystal structure of the monomeric porin OmpG. J. Mol. Biol. 360, 750–759 (2006).Article

Subbarao，G.V。和van den Berg，B。单体孔蛋白OmpG的晶体结构。J、 分子生物学。360750-759（2006）。文章

CAS

中科院

PubMed

PubMed

Google Scholar

谷歌学者

Huang, C. & Mason, J. T. Geometric packing constraints in egg phosphatidylcholine vesicles. Proc. Natl. Acad. Sci. U. S. A. 75, 308–310 (1978).Article

Huang，C。＆Mason，J.T。鸡蛋磷脂酰胆碱囊泡中的几何堆积约束。程序。纳特尔。阿卡德。科学。U、 《美国判例汇编》75308-310（1978）。文章

ADS

广告

CAS

中科院

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Zasadzinski, J. A., Viswanathan, R., Madsen, L., Garnaes, J. & Schwartz, D. K. Langmuir-Blodgett films. Science (New York NY) 263, 1726–1733 (1994).Article

扎萨津斯基（Zasadzinski），维斯瓦纳坦（Viswanathan），马德森（Madsen），加奈斯（Garnaes），J。和施瓦茨（Schwartz），D.K。朗缪尔（Langmuir-Blodgett）电影公司。《科学》（纽约州纽约市）2631726-1733（1994）。文章

ADS

广告

CAS

中科院

Google Scholar

谷歌学者

Tabaei, S. R., Jackman, J. A., Kim, S. O., Zhdanov, V. P. & Cho, N. J. Solvent-assisted lipid self-assembly at hydrophilic surfaces: factors influencing the formation of supported membranes. Langmuir 31, 3125–3134 (2015).Article

Tabaei，S.R.，Jackman，J.A.，Kim，S.O.，Zhdanov，V.P。＆Cho，N.J。亲水表面的溶剂辅助脂质自组装：影响支撑膜形成的因素。朗缪尔313125–3134（2015）。文章

CAS

中科院

PubMed

PubMed

Google Scholar

谷歌学者

Köster, S. et al. Crystal structure of listeriolysin O reveals molecular details of oligomerization and pore formation. Nat. Commun. 5, 3690 (2014).Article

Köster，S。等人。李斯特菌溶血素O的晶体结构揭示了低聚和孔形成的分子细节。国家公社。53690（2014）。文章

ADS

广告

PubMed

PubMed

Google Scholar

谷歌学者

Sonnen, A. F. P., Plitzko, J. M. & Gilbert, R. J. C. Incomplete pneumolysin oligomers form membrane pores. Open Biol. 4, 140044–140044 (2014).Article

Sonnen，A.F.P.，Plitzko，J.M。和Gilbert，R.J.C。不完全的溶血素低聚物形成膜孔。打开Biol。4140044–140044（2014）。文章

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Mulvihill, E., van Pee, K., Mari, S. A., Muller, D. J. & Yildiz, O. Directly observing the lipid-dependent self-assembly and pore-forming mechanism of the Cytolytic Toxin Listeriolysin O. Nano Lett. 15, 6965–6973 (2015).Article

Mulvihill，E.，van Pee，K.，Mari，S.A.，Muller，D.J。＆Yildiz，O。直接观察溶细胞毒素李斯特菌溶血素O.Nano Lett的脂质依赖性自组装和成孔机制。156965-6973（2015）。文章

ADS

广告

PubMed

PubMed

Google Scholar

谷歌学者

van Pee, K., Mulvihill, E., Muller, D. J. & Yildiz, O. Unraveling the pore-forming steps of Pneumolysin from Streptococcus pneumoniae. Nano Lett. 16, 7915–7924 (2016).Article

van Pee，K.，Mulvihill，E.，Muller，D.J。＆Yildiz，O。揭示肺炎链球菌溶血素的成孔步骤。纳诺莱特。167915-7924（2016）。文章

ADS

广告

PubMed

PubMed

Google Scholar

谷歌学者

van Pee, K. et al. CryoEM structures of membrane pore and prepore complex reveal cytolytic mechanism of Pneumolysin. Elife 6, e23644 (2017).Article

van Pee，K。等人。膜孔和预孔复合物的冷冻电镜结构揭示了溶血素的细胞溶解机制。Elife 6，e23644（2017）。文章

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Ilangumaran Ponmalar, I., Sarangi, N. K., Basu, J. K. & Ayappa, K. G. Pore forming protein induced biomembrane reorganization and dynamics: a focused review. Front. Mol. Biosci. 8, 737561 (2021).Article

Ilangumaran-Ponmalar，I.，Sarangi，N.K.，Basu，J.K。＆Ayappa，K.G。成孔蛋白诱导的生物膜重组和动力学：重点综述。正面。。8737561（2021）。文章

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Parsons, E. S. et al. Single-molecule kinetics of pore assembly by the membrane attack complex. Nat. Commun. 10, 2066 (2019).Article

Parsons，E.S.等人，《膜攻击复合物组装孔的单分子动力学》。国家公社。102066（2019）。文章

ADS

广告

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Wang, Y., Zhao, Y., Bollas, A., Wang, Y. & Au, K. F. Nanopore sequencing technology, bioinformatics and applications. Nat. Biotechnol. 39, 1348–1365 (2021).Article

Wang，Y.，Zhao，Y.，Bollas，A.，Wang，Y。＆Au，K.F。纳米孔测序技术，生物信息学和应用。美国国家生物技术公司。391348-1365（2021）。文章

CAS

中科院

PubMed

PubMed

PubMed Central

公共医学中心

Google Scholar

谷歌学者

Niehuis, E., Heller, T., Feld, H., Benninghoven, A. & Vacuum, A. Design and performance of a reflectron based time-of‐flight secondary ion mass spectrometer with electrodynamic primary ion mass separation. J. Vacuum Sci. Technol. Surfaces Films 5, 1243–1246 (1987).Benninghoven, A., Vickerman, J.

Niehuis，E.，Heller，T.，Feld，H.，Benninghoven，A。＆Vacuum，A。基于反射子的飞行时间二次离子质谱仪的设计和性能，具有电动一次离子质量分离。J、 真空科学。技术。表面薄膜51243-1246（1987）。Benninghoven，A.，Vickerman，J。

C. & Briggs, D. TOF-SIMS: Materials Analysis by Mass Spectrometry.Benninghoven, A. 64–89 (Springer, 1983).Wilm, M. & Mann, M. Analytical properties of the nanoelectrospray ion source. Anal. Chem. 68, 1–8 (1996).Article .

C、 ＆Briggs，D。TOF-SIMS：通过质谱法进行材料分析。Benninghoven，A.64-89（Springer，1983）。Wilm，M。＆Mann，M。纳米电喷雾离子源的分析性质。肛门。化学。68，1-8（1996）。文章。

CAS

中科院

PubMed

PubMed

Google Scholar

谷歌学者

Yilmaz, N. & Kobayashi, T. Visualization of lipid membrane reorganization induced by a pore-forming toxin using high-speed atomic force microscopy. ACS Nano 9, 7960–7967 (2015).Article

Yilmaz，N。＆Kobayashi，T。使用高速原子力显微镜观察由成孔毒素诱导的脂质膜重组。ACS Nano 97960–7967（2015）。文章

CAS

中科院

PubMed

PubMed

Google Scholar

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Download referencesAcknowledgementsThe first experiments leading to the electrospray theory and the construction of the first nanoelectrospray were carried out in the laboratory of Prof. A. Benninghoven at the University of Münster, Germany. The experimental confirmation that the nanoelectrospray works as a molecular beam device for molecules of almost unlimited size was obtained in the laboratory of Matthias Mann at the European Molecular Biology Laboratory (EMBL) in Heidelberg, Germany.

下载参考文献致谢导致电喷雾理论的第一个实验和第一个纳米电喷雾的构建是在德国明斯特大学A.Benninghoven教授的实验室进行的。在德国海德堡欧洲分子生物学实验室（EMBL）的马蒂亚斯·曼实验室（MatthiasMann）获得了纳米电喷雾作为几乎无限大小分子的分子束装置的实验证实。

The membrane protein experiments were carried out in the laboratory of Prof. W. Kühlbrandt at the Max Planck Institute of Biophysics, Frankfurt, Germany. I am grateful to Prof. Kühlbrandt for providing the necessary support to carry out all these experiments. I thank Dr. Oezkan Yildiz and Dr. Katharina von Pee for their intellectual and material support.

膜蛋白实验是在德国法兰克福马克斯·普朗克生物物理研究所W.Kühlbrandt教授的实验室进行的。我感谢Kühlbrandt教授为进行所有这些实验提供了必要的支持。我感谢Oezkan Yildiz博士和Katharina von Pee博士的智力和物质支持。

Without the help of Deryck Mills with the electron microscope and with technical questions, the project would never have been completed.FundingThe work was supported by Science Foundation Ireland, SFI grant 07/SK/B1184c. Author informationAuthors and AffiliationsPhysics Institute of the University Münster, Surface Science, Münster, GermanyMatthias WilmEuropean Molecular Biology Laboratory (EMBL), Heidelberg, GermanyMatthias WilmMax Planck Institute for Biophysics, Frankfurt, GermanyMatthias WilmConway Institute, University College Dublin, Belfield, Dublin 4, IrelandMatthias WilmAuthorsMatthias WilmView author publicationsYou can also search for this author in.

如果没有德瑞克·米尔斯（DeryckMills）对电子显微镜和技术问题的帮助，该项目将永远不会完成。资助这项工作得到了爱尔兰科学基金会SFI grant 07/SK/B1184c的支持。作者信息作者和附属机构明斯特大学物理研究所，明斯特表面科学，德国马蒂亚斯·威尔梅欧分子生物学实验室（EMBL），海德堡，德国马蒂亚斯·威尔马克斯·普朗克生物物理学研究所，法兰克福，德国马蒂亚斯·威尔姆康韦研究所，都柏林大学学院，贝尔菲尔德，都柏林4，IrelandMatthias WilmAuthorsMatthias WilmView作者出版物您也可以在中搜索这位作者。

PubMed Google ScholarContributionsThe author M. W. did all the experiments and cited calculations relating to the electrospray theory and wrote the article.Corresponding authorCorrespondence to

PubMed谷歌学术贡献作者M.W.做了所有实验，并引用了与电喷雾理论有关的计算，并撰写了这篇文章。对应作者对应

Matthias Wilm.Ethics declarations

马蒂亚斯·威尔姆。道德宣言

Competing interests

相互竞争的利益

The author holds an European patent for the preparation method for bioactive membranes, EP3711848.

作者拥有生物活性膜制备方法EP3711848的欧洲专利。

Additional informationPublisher’s noteSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.Electronic supplementary materialBelow is the link to the electronic supplementary material.Supplementary Material 1Supplementary Material 2Rights and permissions.

。电子补充材料流是指向电子补充材料的链接。补充材料1补充材料2权利和许可。

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.

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To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/..

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Reprints and permissionsAbout this articleCite this articleWilm, M. Nanoelectrospray based synthesis of large, transportable membranes with integrated membrane proteins.

转载和许可本文引用本文Wilm，M。基于纳米电喷雾的大型可运输膜与整合膜蛋白的合成。

Sci Rep 14, 25192 (2024). https://doi.org/10.1038/s41598-024-76797-wDownload citationReceived: 11 July 2024Accepted: 16 October 2024Published: 24 October 2024DOI: https://doi.org/10.1038/s41598-024-76797-wShare this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

科学报告1425192（2024）。https://doi.org/10.1038/s41598-024-76797-wDownload引文接收日期：2024年7月11日接受日期：2024年10月16日发布日期：2024年10月24日OI：https://doi.org/10.1038/s41598-024-76797-wShare本文与您共享以下链接的任何人都可以阅读此内容：获取可共享链接对不起，本文目前没有可共享的链接。复制到剪贴板。

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