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High-throughput Enzyme Nanopatterning


High-throughput and large-scale patterning of enzymes with sub-10-nm resolution, the size range of individual protein molecules, is crucial for propelling the advancement in a variety of areas, from the development of chip-based biomolecular nano-devices to the molecular-level studies of cell biology. Despite recent developments in bio-nanofabrication technology, combining 10-nm resolution with high-throughput and large-scale patterning of enzymes is still an open challenge. Here, we demonstrate a high resolution and high-throughput patterning method to generate enzyme nanopatterns with sub-10 nm resolution by using thermochemical scanning probe lithography (tc-SPL). First, tc-SPL is used to generate amine patterns on a methacrylate copolymer film. Thermolysin enzymes functionalized with sulfonate-containing fluorescent labels (Alexa-488) are then directly immobilized onto the amine patterns through electrostatic interaction. Enzyme patterns with sub-10 nm line width are obtained as evidenced by atomic force microscopy (AFM) and fluorescence microscopy. Moreover, we demonstrate large-scale and high throughput (0.13 ×0.1 mm^2 at a throughput of 5.2 ×104 µm^2/h) patterning of enzymes incorporating 10-nm detailed pattern features. This straightforward and high-throughput method of fabricating enzyme nanopatterns will have a significant impact on future bio-nanotechnology applications and molecular-level biological studies. By scaling up using parallel probes, tc-SPL is promising for implementation to scale up the fabrication of nano-biodevices.

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Publication details

The article was accepted on 11 Mar 2019 and first published on 14 Mar 2019

Article type: Paper
DOI: 10.1039/C9FD00025A
Faraday Discuss., 2019, Accepted Manuscript

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    High-throughput Enzyme Nanopatterning

    X. Liu, M. Kumar, A. Calo, E. Albisetti, X. Zheng, K. B. Manning, E. Elacqua, M. Weck, R. Ulijn and E. Riedo, Faraday Discuss., 2019, Accepted Manuscript , DOI: 10.1039/C9FD00025A

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