Printed biogel nanosensors
Inventors
McBeth, Christine • Borchers, Kirsten • Weber, Achim • Zontar, Daniel
Assignees
Fraunhofer Gesellschaft zur Foerderung der Angewandten Forschung eV • Boston University • Fraunhofer USA Inc
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Publication Number
US-11813604-B2
Publication Date
2023-11-14
Expiration Date
Abstract
Disclosed is a biogel nanosensor for detection of an analyte that includes an acryloyl or methacryloyl modified hydrogel and nucleic acid amplification reagents in picoliter or nanoliter volume in the form of microarray. Also disclosed are methods of making the disclosed biogel nanosensor, and methods of using the biogel nanosensors.
Core Innovation
The invention provides a biogel nanosensor for detecting an analyte in a sample. The nanosensor includes an acryloyl or a methacryloyl modified hydrogel on the surface of a substrate and nucleic acid amplification reagents. The hydrogel is crosslinked in picoliter or nanoliter volume on the surface of the substrate in a microarray form of spots, while the nucleic acid amplification reagents are layered in picoliter or nanoliter volume on the crosslinked hydrogel spots.
A key aspect is the microarray spot format in very small volumes, where nucleic acid amplification reagents are combined with and localized on crosslinked hydrogel spots on a substrate. The documented disclosure further specifies that the nucleic acid amplification reagents can comprise DNA or RNA with primers, a polymerase, and a detection element. The detection element can be fluorescent, colorimetric, or electrochemical.
The disclosure also covers preparation of the biogel nanosensor by obtaining an acryloyl hydrogel or a methacryloyl hydrogel, printing the hydrogel in picoliter or nanoliter volume on the surface of a substrate in a microarray in the form of spots, crosslinking the hydrogel on the surface of the substrate, and combining nucleic acid amplification reagents in picoliter or nanoliter volume to the cross-linked hydrogel. Additional elements described include a light guide and/or a heating element, with the heating element triggering nucleic acid amplification after the biogel nanosensor contacts the sample.
Claims Coverage
The partial content includes two independent claims. Across these claims, the inventive core features are: a microarray of crosslinked acryloyl/methacryloyl modified hydrogel spots with nucleic acid amplification reagents layered in picoliter or nanoliter volume, and a preparation method printing and crosslinking the hydrogel microarray followed by combining nucleic acid amplification reagents in similarly small volumes.
Microarray biogel nanosensor with crosslinked acryloyl/methacryloyl hydrogel spots and picoliter/nanoliter nucleic acid amplification layering
A biogel nanosensor for detecting an analyte comprising an acryloyl or a methacryloyl modified hydrogel on a substrate surface, nucleic acid amplification reagents, wherein the hydrogel is crosslinked in picoliter or nanoliter volume on the substrate surface in a microarray form of spots, and the nucleic acid amplification reagents are layered in picoliter or nanoliter volume on the crosslinked hydrogel spots.
Method of preparing a biogel nanosensor by printing acryloyl/methacryloyl hydrogel microarray spots, crosslinking, then combining nucleic acid amplification reagents in picoliter/nanoliter volume
A method of preparing a biogel nanosensor comprising obtaining an acryloyl hydrogel or a methacryloyl hydrogel, printing the hydrogel in picoliter or nanoliter volume on the surface of a substrate in a microarray in the form of spots, crosslinking the hydrogel on the surface of the substrate, and combining nucleic acid amplification reagents in picoliter or nanoliter volume to the cross-linked hydrogel.
Claim coverage in this partial content centers on forming a microarray of crosslinked acryloyl or methacryloyl modified hydrogel spots on a substrate and placing nucleic acid amplification reagents onto those spots in picoliter or nanoliter volumes, including a preparation sequence that prints, crosslinks, and then combines the nucleic acid amplification reagents in picoliter or nanoliter volume.
Stated Advantages
Improved performance including a 100-fold LOD improvement to ~10 copies/reaction [as described in the provided partial content summary].
Documented Applications
Demonstration of inkjet-printed DENV RT-LAMP signal detection [as described in the provided partial content summary].
Multiplexed potential for detecting viral analytes such as DENV, and comparing detection against other viruses including Zika virus, Influenza A/B, Rhinovirus, and SARS-CoV-2 [as described in the provided partial content summary].
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