Synthetic human cell mimic particle for cytometric or coulter device
Inventors
Kim, Jeffrey • LIU, OLIVER • Agresti, Jeremy • Nguyen, Anh Tuan
Assignees
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Publication Number
US-12352679-B2
Publication Date
2025-07-08
Expiration Date
Abstract
Synthetic human cell mimic hydrogel particles and their use in cytometric or coulter device applications are described. The synthetic human cell mimic hydrogel particles described herein are selectively tunable to have at least one optical, volumetric, or capacitance property that is substantially similar to a corresponding optical, volumetric, or capacitance property of the human cell mimic hydrogel particle's natural biological cell counterpart.
Core Innovation
The invention relates to synthetic human cell-mimic hydrogel particles that include a polymer matrix configured to mimic cytometric properties of natural biological cells. The particles are used as cell-like fluorescence and optical scatter standards for detecting cell-surface markers by flow cytometry, including FACS, and for generating measurable fluorescence standard curves.
A core aspect is attaching biomolecules to the hydrogel particle so that the particle comprises a polymer matrix attached to a plurality of different biomolecules, including CD3 and/or CD28. The particles have tunable optical and volumetric properties, including forward scatter (FSC), side scatter (SSC), and fluorescence emission profiles, and the optical-scatter properties can substantially match target human cell counterparts.
The hydrogel particle design is directed to overcome limitations of conventional polystyrene calibration beads by enabling optical properties to be modulated and matched to natural cell counterparts. Optical properties are independently modulated via hydrogel composition, including monomer and co-monomer amounts, surface functionalization, polymerization initiator/crosslinker choices, refractive index tuning, and hydrogel optical tuning with biomolecule or nanoparticle loading.
The document further describes generating standard curves and providing assay controls using these synthetic particles for calibration and detection or sorting standards for cytometric or coulter devices. It also broadens the target definitions beyond traditional cells to include tumor microvesicles/macrovesicles and discusses flow cytometers compatible with hydrogel assays.
Claims Coverage
The claim coverage centers on one independent hydrogel particle claim supported by multiple refinements. The inventive features focus on a polymer matrix attached to biomolecules comprising CD3 and/or CD28, together with optical scatter matching, particle diameter, and polymerized monomer formation.
Hydrogel particle with biomolecules comprising CD3 and/or CD28
A hydrogel particle comprising a polymer matrix attached to a plurality of different biomolecules comprising CD3 and/or CD28.
Optical scatter property substantially matches side scatter (SSC)
The hydrogel particle has an optical-scatter property that substantially matches the side scatter (SSC) of a natural biological cell counterpart.
Optical scatter property substantially similar for forward scatter (FSC)
The hydrogel particle has an optical-scatter property substantially similar to that of the corresponding natural biological cell counterpart for forward scatter (FSC).
Particle diameter range
The hydrogel particle has a diameter ranging from 1–100 μm.
Hydrogel polymer matrix formed by polymerizing a monomer
The hydrogel polymer matrix is formed by polymerizing a monomer.
Overall, the claim coverage centers on hydrogel particles with a polymer matrix attached to biomolecules comprising CD3 and/or CD28, and extends to optical-scatter matching using SSC and FSC metrics, along with particle diameter and polymerized monomer formation.
Stated Advantages
Provides tunable optical properties, including forward scatter (FSC), side scatter (SSC), and fluorescence emission profiles, that can substantially match natural cell counterparts.
Provides synthetic calibration and standard-like cytometric particles for calibration, target-cell detection, sorting, and assay controls.
Documented Applications
Flow cytometry calibration using synthetic human cell-mimic hydrogel particles with optical-scatter matching to natural biological cells.
Target-cell detection and sorting using the hydrogel particles in cytometric contexts described in the document.
Generating standard curves and assay controls in cytometric or coulter devices.
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