Issue 22, 2020

High-throughput roll-to-roll production of polymer biochips for multiplexed DNA detection in point-of-care diagnostics

Abstract

Roll-to-roll UV nanoimprint lithography has superior advantages for high-throughput manufacturing of micro- or nano-structures on flexible polymer foils with various geometries and configurations. Our pilot line provides large-scale structure imprinting for cost-effective polymer biochips (4500 biochips/hour), enabling rapid and multiplexed detections. A complete high-volume process chain of the technology for producing structures like μ-sized, triangular optical out-couplers or capillary channels (width: from 1 μm to 2 mm, height: from 200 nm up to 100 μm) to obtain biochips (width: 25 mm, length: 75 mm, height: 100 μm to 1.5 mm) was described. The imprinting process was performed with custom-developed resins on polymer foils with resin thicknesses ranging between 125–190 μm. The produced chips were tested in a commercial point-of-care diagnostic system for multiplexed DNA analysis of methicillin resistant Staphylococcus aureus (e.g., mecA, mecC gene detections). Specific target DNA capturing was based on hybridisation between surface bound DNA probes and biotinylated targets from the sample. The immobilised biotinylated targets subsequently bind streptavidin–horseradish peroxidase conjugates, which in turn generate light upon incubation with a chemiluminescent substrate. To enhance the light out-coupling thus to improve the system performance, optical structures were integrated into the design. The limits-of-detection of mecA (25 bp) for chips with and without structures were calculated as 0.06 and 0.07 μM, respectively. Further, foil-based chips with fluidic channels were DNA functionalised in our roll-to-roll micro-array spotter following the imprinting. This straightforward approach of sequential imprinting and multiplexed DNA functionalisation on a single foil was also realised for the first time. The corresponding foil-based chips were able to detect mecA gene DNA sequences down to a 0.25 μM concentration.

Graphical abstract: High-throughput roll-to-roll production of polymer biochips for multiplexed DNA detection in point-of-care diagnostics

Supplementary files

Article information

Article type
Paper
Submitted
24 Jul 2020
Accepted
06 Oct 2020
First published
22 Oct 2020
This article is Open Access
Creative Commons BY-NC license

Lab Chip, 2020,20, 4106-4117

High-throughput roll-to-roll production of polymer biochips for multiplexed DNA detection in point-of-care diagnostics

P. Toren, M. Smolka, A. Haase, U. Palfinger, D. Nees, S. Ruttloff, L. Kuna, C. Schaude, S. Jauk, M. Rumpler, B. Hierschlager, I. Katzmayr, M. Sonnleitner, M. W. Thesen, M. Lohse, M. Horn, W. Weigel, M. Strbac, G. Bijelic, S. Hemanth, N. Okulova, J. Kafka, S. Kostler, B. Stadlober and J. Hesse, Lab Chip, 2020, 20, 4106 DOI: 10.1039/D0LC00751J

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