# US 11,306,288 B2 · Methods for preparing therapeutically active cells using microfluidics > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,306,288 B2 - Document ID: US11306288B2 - Status: Granted - Kind: B2 - Application number: 17/192,691 - Filed: 2021-03-04 - Published: 2021-07-08 - Granted: 2022-04-19 - Earliest claimed: 2017-09-01 - Assignees: GPB Scientific, LLC; University of Maryland, Baltimore; The Trustees of Princeton University - Applicants: Gpb Scientific, Inc.; University of Maryland, Baltimore; The Trustees of Princeton University - Inventors: Anthony Ward; Roberto Campos-Gonzalez; Alison Skelley; Khushroo Gandhi; Curt Civin; James C. Sturm; Michael Grisham - Web page: https://patentgrep.com/patent/US11306288B2 ## CPC - C12N 5/0636: {T lymphocytes} - A61K 40/11: T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells - A61K 40/31: Chimeric antigen receptors [CAR] - A61K 40/42: Cancer antigens - A61K 40/4254: {Adhesion molecules, e.g. NRCAM, EpCAM or cadherins} - A61P 35/02: specific for leukemia - B01L 3/502753: {characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation} - B01L 3/502761: {specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules} - C12N 5/0087: {Purging against subsets of blood cells, e.g. purging alloreactive T cells} - B01L 2200/0652: Sorting or classification of particles or molecules - B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions - B01L 2300/0864: comprising only one inlet and multiple receiving wells, e.g. for separation, splitting - B01L 2400/086: using baffles or other fixed flow obstructions - C12N 2531/00: Microcarriers - C12N 2533/54: Collagen; Gelatin - C12N 2533/74: Alginate - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE ## Abstract The present invention is directed to the use of microfluidics in the preparation of cells and compositions for therapeutic uses. ## Claims ### Claim 1 A method for preparing a purified leukocyte composition comprising: a) purifying leukocytes from an apheresis sample, wherein the sample comprises leukocytes and platelets and the leukocytes are separated from the platelets by: i) performing a size-based separation using a microfluidic device configured to separate cells by deterministic lateral displacement to produce an enriched product in which, compared to the sample, the percentage of platelets has decreased and the percentage of leukocytes has increased; ii) in addition to the size-based separation, performing an affinity-based separation by binding one or more types of leukocytes to a carrier that binds to the one or more types of leukocytes with specificity, and then separating the carrier-bound cells from cells not bound to carrier; b) after the purification of step a), activating and expanding the purified leukocytes in culture to produce the purified leukocyte composition. ### Claim 2 (depends on 1) The method of claim 1, wherein the apheresis sample is obtained from a patient and the patient has rheumatoid arthritis, type I diabetes, or multiple sclerosis. ### Claim 3 (depends on 1) The method of claim 1, wherein, after the purified leukocyte composition is prepared, said composition is administered to a patient. ### Claim 4 (depends on 1) The method of claim 1, wherein the platelets in the enriched product of paragraph a)i) are depleted by at least 80% compared to the sample and/or there are no more than 5 platelets per leukocyte in the enriched product. ### Claim 5 (depends on 1) The method of claim 1, further comprising: c) genetically engineering the activated leukocytes. ### Claim 6 (depends on 1) The method of claim 1, wherein the apheresis sample is obtained from a patient and the patient has cancer. ### Claim 7 (depends on 1) The method of claim 1, wherein the leukocytes in the purified leukocyte composition are selected from the group consisting of: B-cells, NK-cells, and monocytes. ### Claim 8 (depends on 1) The method of claim 1, wherein the leukocytes in the purified leukocyte composition are granulocytes. ### Claim 9 (depends on 1) The method of claim 1, wherein the leukocytes in the purified leukocyte composition are selected from the group consisting of: central memory T cells; helper T cells and regulatory T cells. ### Claim 10 (depends on 1) The method of claim 1, wherein the sample was obtained by leukapheresis. ### Claim 11 (depends on 1) The method of claim 1, wherein the cells of the purified leukocyte composition are transferred into a pharmaceutical composition and are not frozen prior to transfer. ### Claim 12 (depends on 5) The method of claim 5, wherein the apheresis sample is obtained from a patient and the patient has cancer. ### Claim 13 (depends on 12) The method of claim 12, wherein the leukocytes in the purified leukocyte composition are selected from the group consisting of: B-cells, NK-cells, and monocytes. ### Claim 14 (depends on 12) The method of claim 12, wherein the leukocytes in the purified leukocyte composition are granulocytes. ### Claim 15 (depends on 12) The method of claim 12, wherein the leukocytes in the purified leukocyte composition are selected from the group consisting of: central memory T cells; helper T cells and regulatory T cells. ### Claim 16 (depends on 12) The method of claim 12, wherein the sample was obtained by leukapheresis. ### Claim 17 (depends on 12) The method of claim 12, wherein the cells of the purified leukocyte composition are transferred into a pharmaceutical composition and are not frozen prior to transfer.