Methods and related aspects for multiplexed analyte detection using sequential magnetic particle elution
Inventors
Wang, Tza-Huei Jeff • TRICK, Alexander Y. • Chen, Fan-En • Chen, Liben
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Assignees
MemberJohns Hopkins UniversityJohns Hopkins UniversityFounded in 1876, Johns Hopkins University is recognized as the first research university in the United States. It advances interdisciplinary education, high-impact research, and global outreach, supporting knowledge translation, technological innovation, and community partnerships. The university fosters academic excellence, innovation incubation, outreach, and inclusion across multiple campuses in Baltimore, integrating into the city's social, economic, and cultural life.
Founded in 1876, Johns Hopkins University is recognized as the first research university in the United States. It advances interdisciplinary education, high-impact research, and global outreach, supporting knowledge translation, technological innovation, and community partnerships. The university fosters academic excellence, innovation incubation, outreach, and inclusion across multiple campuses in Baltimore, integrating into the city's social, economic, and cultural life.
Publication Number
US-12697618-B2
Publication Date
2026-08-04
Expiration Date
Abstract
Provided herein are magnetofluidic cartridges of use in a wide variety of sample analysis applications, including nucleic acid amplification assays. The magnetofluidic cartridges include sample inlet wells and sample analysis wells for performing controlled serial elution techniques that enables execution of extraction/purification and splitting of analytes for multiplex detection via magnetic actuation only. Related magnetofluidic devices, kits, and methods are also provided.
Core Innovation
A method detects multiple biomolecule species in a magnetofluidic cartridge using magnetic particles and magnetic actuation only. The method includes contacting a sample inlet with a plurality of magnetic particles so that a first biomolecule species and a second biomolecule species bind to the magnetic particles, then moving the bound biomolecule species through sequential fluid communication into separate sample analysis wells using at least one magnet.
In each sample analysis well, a portion of the corresponding bound biomolecule species is eluted to produce an unbound biomolecule species, while remainder bound material is moved onward to the next analysis well. The method performs detection of unbound biomolecule species, or derivatives thereof or both, in the first sample analysis well and detection of the second unbound biomolecule species, or derivatives thereof or both, in the second sample analysis well.
The magnetofluidic cartridge architecture includes a sample inlet well, a first sample analysis well, and a second sample analysis well that are fluidly coupled in a series such that the first sample analysis well fluidly communicates with the sample inlet well and the second sample analysis well fluidly communicates with the first sample analysis well. The cartridge includes a layered construction, cartridge reagent differentiation, and tuning of elution by selecting one or more elution conditions, including temperature, duration, buffer composition, reaction mixture composition, and pH level.
Claims Coverage
The provided claims content identifies one independent claim and no other explicitly listed independent claims. The main inventive features cover sequential magnetically actuated transport through multiple analysis wells, partial elution into unbound biomolecule species in each well, and detection of unbound biomolecule species in corresponding wells.
Sequential magnetically actuated transport and partial elution for multiple biomolecule species detection
A method detecting a first biomolecule species and a second biomolecule species using a magnetofluidic cartridge that binds both species to a plurality of magnetic particles at a sample inlet, moves the bound species to a first sample analysis well using at least one magnet, produces a first unbound biomolecule species in the first sample analysis well by eluting a portion of the first bound biomolecule species, moves the remainder of the first bound biomolecule species and the second bound biomolecule species to a second sample analysis well, produces a second unbound biomolecule species in the second sample analysis well by eluting a portion of the second bound biomolecule species, and detects the unbound biomolecule species, or derivatives thereof or both, in the respective wells.
Washing prior to elution
The method further includes washing first and second bound biomolecule species on magnetic particles before eluting the first or the second bound biomolecule species.
Layered magnetofluidic cartridge architecture with sealable fluid channel and different processing reagents
A magnetofluidic cartridge configured with top and bottom layers defining multiple wells including a sample inlet well and a first and second sample analysis wells, a spacer-defined fluid channel communicating with the wells via a port sealable in a closed position, and magnetic particles in the sample inlet well and a different processing reagent in each sample analysis well.
Tunable elution conditions for sequential partial elution
Tuning one or more elution conditions to elute portions of first and second bound biomolecule species from a plurality of magnetic particles.
Elution conditions defined by selected parameters
Elution conditions defined by selected parameters including temperature, duration, buffer composition, reaction mixture composition, and pH level.
Biomolecule species corresponding to different genomic DNA loci or variants
The first and second biomolecule species correspond to different loci of genomic DNA, or different genetic locus variants.
Across the provided claims, the inventive concept is sequential magnetically actuated movement of magnetic-particle-bound biomolecule species through multiple analysis wells, where portions of bound species are eluted to generate unbound biomolecule species in each well followed by detection in the corresponding well. Coverage also includes optional washing, layered cartridge architecture with sealable fluid communication and different processing reagents, and elution tuned using specified elution-condition parameters, including genomic DNA loci or variant-based biomolecule species.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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