# US 12,216,116 B2 · Magnetic separation of biological entities from fluid sample > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,216,116 B2 - Document ID: US12216116B2 - Status: Granted - Kind: B2 - Application number: 17/579,528 - Filed: 2022-01-19 - Published: 2022-05-12 - Granted: 2025-02-04 - Earliest claimed: 2018-07-20 - Assignees: Cornell University - Applicants: Cornell University - Inventors: Sasank Vemulapati; David Erickson - Web page: https://patentgrep.com/patent/US12216116B2 ## CPC - G01N 33/54326: {Magnetic particles} - B03C 1/01: by addition of magnetic adjuvants - B03C 1/288: {disposed at the outer circumference of a recipient} - G01N 1/4077: {by other techniques involving separation of suspended solids} - G01N 33/56966: {Animal cells} - G01N 33/80: involving blood groups or blood types {or red blood cells (white blood cells G01N33/56972)} - G01N 35/0098: {involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation (magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C)} - B03C 2201/18: Magnetic separation whereby the particles are suspended in a liquid - B03C 2201/26: for use in medical or biological applications - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - B03C: MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS (separating isotopes B01D59/00; combinations of magnetic or electrostatic separation with separation of solids by other means B03B, B07B) ## Abstract The present disclosure relates to, inter alia, devices, systems, and methods for use in the magnetic separation of biological entities from fluid samples. This device includes a magnetic separation chamber configured to receive a fluid sample for magnetic separation, where the magnetic separation chamber includes at least two magnets mounted on the surface or in the wall of the magnetic separation chamber. The device also includes a force provider configured to move the magnetic separation chamber in a side-to-side motion to mix and/or magnetize the fluid sample. In one embodiment, the magnetic separation chamber is in a form of a sleeve and comprises a substantially central channel for loading a vessel containing the fluid sample therein. The systems and methods of the present disclosure involve the use of this device to separate biological entities from fluid samples. ## Claims ### Claim 1 A system for use in magnetic separation of a target biological entity from a blood sample, said system comprising: a vessel comprising a tube or a container for containing the blood sample during operation; a plurality of magnetic beads each functionalized to bind to the target biological entity; an aggregation agent configured to induce clumping of a plurality of target biological entities to form aggregates of the target biological entities, wherein the aggregation agent has a concentration of about 3 mg/mL to about 4 mg/mL; and a magnetic source configured to generate a magnetic field within the vessel to separate the plurality of magnetic beads having bound aggregates of the target biological entities from the blood sample, such that the target biological entities are substantially captured and depleted to separate greater than 99.9% of the target biological entity. ### Claim 2 (depends on 1) The system according to claim 1, wherein said aggregation agent is selected from the group consisting of a polyclonal antibody, monoclonal antibody, a protein, dextran, polyvinylpyrollidone, polyoxyethylene, fibrinogen, immunoglobulin IgM and IgG, calcium chloride, and combinations thereof. ### Claim 3 (depends on 1) The system according to claim 1, wherein the magnetic beads are each functionalized with an anti-blood cell antibody that binds to red blood cells, white blood cells, platelets, or any combinations thereof, specifically or non-specifically. ### Claim 4 (depends on 1) The system according to claim 1, wherein the aggregation agent is an antibody that binds to red blood cells, white blood cells, platelets, or any combinations thereof, specifically or non-specifically. ### Claim 5 (depends on 1) The system according to claim 1, wherein the magnetic beads are conjugated to the aggregation agent and the aggregation agent is an antibody that binds to red blood cells, white blood cells, platelets, or any combinations thereof specifically or non-specifically. ### Claim 6 (depends on 1) The system according to claim 1, wherein the aggregation agent is an antibody that binds to a surface marker of red blood cells selecting from the group consisting of CD235a and TER119. ### Claim 7 (depends on 1) The system according to claim 1, wherein the target biological entity is selected from the group consisting of red blood cells, white blood cells, platelets, cell associated contaminants, waste products, and excess reagents contained in the blood sample. ### Claim 8 (depends on 1) The system according to claim 1, further comprising a collection component for collecting a supernatant produced after capturing and separating the target biological entity from the blood sample. ### Claim 9 (depends on 1) The system according to claim 1 wherein the magnetic separation of target biological entities from blood is achieved without any centrifugation or filtration steps in the system. ### Claim 10 (depends on 1) The system according to claim 1, wherein the vessel comprises magnetic beads and dried aggregation agent. ### Claim 11 (depends on 1) The system according to claim 1, wherein the system is configured to at least substantially homogenously distribute the magnetic beads and the aggregation agent in the blood sample. ### Claim 12 (depends on 1) The system according to claim 1, wherein the magnetic source comprises two magnets on either side of the vessel configured to move in a side-to-side motion with respect to the blood sample to achieve at least substantially homogenous distribution. ### Claim 13 (depends on 1) The system according to claim 1, further comprising a force provider to provide a sufficient number of strokes to at least substantially homogenously distribute the functionalized magnetic beads and aggregation agent in the blood sample within no more than 90 seconds. ### Claim 14 (depends on 1) The system according to claim 1, wherein the system is configured to separate greater than 99.9% of target biological entity within 2 minutes. ### Claim 15 (depends on 13) The system according to claim 13, wherein the force provider is configured to at least substantially homogeneously distribute the functionalized magnetic beads and aggregation agent in the blood sample within no more than 90 seconds and the magnetic field is configured to concentrate the magnetic beads and the target biological entity on one side of the vessel. ### Claim 16 (depends on 1) The system according to claim 1, further comprising a housing unit to house the vessel, a force provider, and one or more electronics components configured to operate the force provider and automate the system to separate the target biological entity within a predetermined time period. ### Claim 17 (depends on 1) The system according to claim 1, further comprising a collection component configured to collect a supernatant product after the separation of the target biological entity from the blood sample, wherein the supernatant product has an average purity of greater than 99.9% and a recovery rate of greater than 90%. ### Claim 18 (depends on 1) The system according to claim 1, wherein the generated magnetic field is configured to homogenously distribute the functionalized magnetic beads among the target biological entity during the laterally side-to-side motion and concentrate the magnetic beads and the target biological entity on one side of the vessel after the laterally side-to-side motion. ### Claim 19 (depends on 1) The system according to claim 1, wherein each functionalized magnetic bead has a plurality of functional groups, and each functional group is configured to capture a plurality of aggregated target biological entities. ### Claim 20 (depends on 1) The system according to claim 1, wherein each magnetic bead is conjugated with a plurality of aggregation agents, each conjugated aggregation agent is configured to capture a plurality of aggregated target biological entities grouped by a plurality of unconjugated aggregation agents. ### Claim 21 (depends on 1) The system according to claim 1, further comprising a force provider configured to move the magnetic field in a specific direction with respect to the sample to create a mixing effect comprising lateral mixing and gravitational mixing within the sample and to separate the plurality of magnetic beads and aggregates of the target biological entities from the blood sample to yield a supernatant with greater than 99.9% of the target biological entities removed without using a centrifuge in the system.