Raman spectroscopy based assay for both low and high abundant biomolecules in a biological fluid sample
Inventors
Fournier, David • Pandey, Rishikesh
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11686684-B2
Publication Date
2023-06-27
Expiration Date
Abstract
A system and method for assaying high and low abundant biomolecules within a biological fluid sample is provided. The method includes: a) placing a biological fluid sample in contact with a first nanostructure surface; b) interrogating the sample with a light source, the sample in contact with the first nanostructure surface, the interrogation using a SERS technique; c) detecting an enhanced Raman scattering from at least one high abundant biomolecule type and producing first signals representative thereof; d) placing the sample in contact with a second nanostructure surface having a targeting agent that targets a low abundant biomolecule; e) interrogating the sample with the light source using the SERS technique; f) detecting the enhanced Raman scattering from the low abundant biomolecules and producing second signals representative thereof; and g) assaying the biological fluid sample using the first signals and the second signals.
Core Innovation
The invention provides an assay for simultaneously assaying one or more high abundant biomolecules and one or more low abundant biomolecules within a biological fluid sample using surface enhanced Raman spectroscopic (SERS) techniques on nanostructure surfaces. A biological fluid sample is placed in contact with a first nanostructure surface, and the sample is interrogated with a light source such that enhanced Raman scattering from at least one high abundant biomolecule adsorbed on the first nanostructure surface is produced and detected. First signals representative of the enhanced Raman scattering from the high abundant biomolecules are generated as part of producing an assay outcome.
The assay further includes placing the biological fluid sample in contact with at least one second nanostructure surface that is functionalized with at least one targeting agent configured to target at least one low abundant biomolecule. The biological fluid sample is interrogated with the light source while in contact with the second nanostructure surface, and the interrogation is configured to produce enhanced Raman scattering from at least one low abundant biomolecule captured by the targeting agent functionalized on the second nanostructure surface. Second signals representative of the enhanced Raman scattering from the low abundant biomolecules are detected and then used together with the first signals to assay the biological fluid sample.
The disclosed approach addresses the problem of assaying both high-abundance and low-abundance biomolecules from the same biological fluid sample in a way that combines SERS detection across two different nanostructure interrogation phases. The method and system separate an initial interrogation of high abundant biomolecules on a nanostructure surface into an untargeted configuration and a subsequent interrogation of low abundant biomolecules on a targeting-agent functionalized nanostructure surface. Signals from both interrogation phases are combined for the assay using presence determination and/or quantity and concentration determination, as described in the partial content.
Claims Coverage
The independent claims are clm-00001, clm-00010, and clm-00011, covering a two-part SERS assay with sequential interrogation of nanostructure surfaces and combination of detected signals for an overall assay result. The inventive features focus on three inventive features: two nanostructure surfaces with different roles, SERS interrogation and detection to generate first and second signals, and control/system architecture for producing the assay outcome using the combined signals.
Sequential SERS interrogation of a first nanostructure surface for high abundant biomolecules then a second targeting-agent nanostructure surface for low abundant biomolecules
Placing a biological fluid sample in contact with a first nanostructure surface and interrogating the sample with SERS to produce enhanced Raman scattering from at least one high abundant biomolecule adsorbed on the first nanostructure surface, detecting first signals representative thereof, then placing the sample in contact with at least one second nanostructure surface functionalized with at least one targeting agent configured to target at least one low abundant biomolecule, interrogating using SERS to produce enhanced Raman scattering from low abundant biomolecules captured by the targeting agent, detecting second signals representative thereof, and assaying the biological fluid sample using the first signals and the second signals.
First nanostructure surface free of biomolecule targeting agents for high abundant biomolecules and second targeting-agent nanostructure surface for low abundant biomolecules
Placing a biological fluid sample in contact with a first nanostructure surface free of any biomolecule targeting agents and interrogating with SERS configured to produce enhanced Raman scattering from at least one type of high abundant biomolecule adsorbed on the first nanostructure surface and detecting first signals representative thereof, then placing the sample in contact with at least one second nanostructure surface functionalized with at least one targeting agent configured to target at least one type of low abundant biomolecule, interrogating to produce Raman scattering from the low abundant biomolecule captured by the targeting agent and detecting second signals representative thereof, and assaying the biological fluid sample using the first signals and the second signals.
System controlling SERS interrogation on an untargeted first nanostructure surface and a targeting-agent second nanostructure surface and assaying using combined signals
Providing a first nanostructure surface free of biomolecular targeting agents and at least one second nanostructure surface functionalized with at least one targeting agent configured to target at least one type of low abundant biomolecule within the biological fluid sample, and a light source, light detector, and system controller with memory storing instructions that cause a processor to control sequential SERS interrogation of the sample on the first surface to produce enhanced Raman scattering from at least one type of high abundant biomolecule and produce first signals, control interrogation on the second surface to produce enhanced Raman scattering from at least one type of low abundant biomolecule and produce second signals, and assay the biological fluid sample using the first signals and the second signals.
Across clm-00001, clm-00010, and clm-00011, the claims cover a SERS-based assay that generates first signals from enhanced Raman scattering of high abundant biomolecules on a first nanostructure surface and second signals from enhanced Raman scattering of targeting-agent-captured low abundant biomolecules on a second nanostructure surface, with an overall assay result determined using the combined first and second signals.
Stated Advantages
Enables assaying both high-abundance and low-abundance biomolecules from the same biological fluid sample using SERS.
Supports presence determination and quantity/concentration determination by combining signals from the two SERS interrogation phases.
Immune to photobleaching.
Improves sensitivity compared with ELISA-like approaches, including detection down to sub-femtomolar ranges as asserted in the partial content.
Documented Applications
Assaying one or more high abundant biomolecules and one or more low abundant biomolecules in a biological fluid sample such as serum, plasma, saliva, urine, or blood.
Presence determination and quantity/concentration determination for high-abundant and low-abundant biomolecules using combined SERS signals from two nanostructure interrogation phases.
Comparison/positioning of the SERS-based assay against ELISA-like approaches, as described in the partial content.
Interested in licensing this patent?