# US 12,110,505 B2 · Method for preparing a leukocyte preparation and leukocyte preparation > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,110,505 B2 - Document ID: US12110505B2 - Status: Granted - Kind: B2 - Application number: 16/506,462 - Filed: 2019-07-09 - Published: 2020-01-02 - Granted: 2024-10-08 - Earliest claimed: 2012-06-08 - Assignees: Artcline GmbH - Applicants: Artcline GmbH; Fraunhofer-Gesellschaft Zur FöRderung Der Angewandten Forschung E.V. - Inventors: Jens Altrichter; Steffen Mitzner; Antje Schwarz; Kathleen Selleng; Fanny Doss; Stephanie Koch - Web page: https://patentgrep.com/patent/US12110505B2 ## CPC - A61K 35/16: Blood plasma; Blood serum (umbilical cord blood A61K35/51) - C12N 5/0636: {T lymphocytes} - 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) - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract A method for preparing a leukocyte preparation from a leukocyte fraction and a correspondingly prepared or obtainable leukocyte preparation and its use. The methods include: a) sedimenting the leukocyte fraction removing leukocyte supernatant from the sediment, wherein the leukocyte supernatant is collected or remains in a leukocyte container, b) sedimenting the leukocytes in the leukocyte container, c) washing the sediment in the leukocyte container with a saline solution, and d) resuspending the sediment in the leukocyte container in a storage solution. ## Claims ### Claim 1 A leukocyte preparation comprising: leukocytes; erythrocytes; and thrombocytes; wherein the leukocyte preparation is produced according to a method comprising: obtaining a leukocyte fraction from whole blood; sedimenting the leukocyte fraction until a dividing line is formed between sedimented erythrocytes and a leukocyte supernatant, removing the leukocyte supernatant from the sediment, wherein the leukocyte supernatant is collected or remains in a leukocyte container; sedimenting the leukocytes in the leukocyte container, wherein a low-leukocyte supernatant is formed, and removing the low-leukocyte supernatant from the leukocyte container; washing the sediment in the leukocyte container with a saline solution, wherein the sedimented cells are resuspended in the saline solution, the solution is sedimented and a resulting supernatant is removed; resuspending the sediment in the leukocyte container in a storage solution; and storing the solution from the resuspending in a gas-permeable bag at a temperature between 20° C. and 25° C. for at least 48 hours; wherein the method modifies the leukocyte preparation to create a leukocyte preparation that is storable at a temperature between 2° C. and 38° C. for a period of 48 hours to 168 hours without substantial loss of function and respiratory burst and phagocytosis are more than 90 percent of a fresh version of the leukocyte preparation. ### Claim 2 (depends on 1) The leukocyte preparation according to claim 1, wherein: with respect to the whole blood, erythrocytes are reduced by more than 90%, thrombocytes are reduced by more than 90%, and leukocytes are reduced by not more than 60%. ### Claim 3 (depends on 1) The leukocyte preparation according to claim 1, wherein: the sedimenting of the leukocyte fraction comprises sedimenting the leukocyte fraction at an acceleration from 0.5 g to 1000 g; or the sedimenting of the leukocyte fraction comprises sedimenting the leukocyte fraction with the addition of a sedimentation accelerator. ### Claim 4 (depends on 3) The leukocyte preparation according to claim 3, wherein the sedimentation accelerator is selected from the group consisting of hydroxyethyl starch, dextran, Ficoll, Percoll and gelatin. ### Claim 5 (depends on 1) The leukocyte preparation according to claim 1, wherein: a duration of the sedimenting of the leukocyte fraction is between 5 and 120 minutes; or the sedimenting of the leukocytes and/or the sedimenting of the solution is carried out by centrifugation. ### Claim 6 (depends on 5) The leukocyte preparation according to claim 5, wherein the centrifugation is at an acceleration from 50 to 10000 g. ### Claim 7 (depends on 1) The leukocyte preparation according to claim 1, wherein the washing of the sediment is repeated one or more times and is processed by one or more of centrifugation, filtration, and filtration with a physiological solution containing 50 to 200 mM sodium chloride. ### Claim 8 (depends on 7) The leukocyte preparation according to claim 7, wherein the physiological solution comprises at least one of a second salt and a buffer. ### Claim 9 (depends on 1) The leukocyte preparation according to claim 1, wherein the gas-permeable bag is a gas-permeable thrombocyte storage bag. ### Claim 10 (depends on 1) The leukocyte preparation according to claim 1, wherein the storage solution is selected from at least one of a blood group-identical blood plasma and an aqueous solution containing blood group-identical plasma. ### Claim 11 (depends on 1) The leukocyte preparation according to claim 1, wherein: the method is carried out aseptically in a closed system; or the method is carried out in a system of tubing and bags. ### Claim 12 (depends on 1) The leukocyte preparation according to claim 1, further comprising adding at least one additive selected from the group consisting of pH-stabilizing substances, cytokines, growth factors, blood group-identical blood plasma, albumin, antioxidants, ascorbic acid, glutathione, cysteine, and methionine. ### Claim 13 (depends on 1) The leukocyte preparation according to claim 1, wherein: before, during or after any of the sedimenting of the leukocyte fraction, the sedimenting of the leukocytes, the washing of the sediment, the resuspending of the sediment, and the storing of the solution, multiple preparations are pooled; before or after at least one of the sedimenting of the leukocyte fraction, the sedimenting of the leukocytes, the washing of the sediment, the resuspending of the sediment, and the storing of the solution or immediately before use, an irradiation takes place; or a portion is transferred from one bag to another by at least one of a pressure difference generated manually, a pressure difference generated by a machine, and gravity. ### Claim 14 (depends on 1) The leukocyte preparation of claim 1, further comprising PAGGS-M. ### Claim 15 (depends on 1) The leukocyte preparation of claim 1, wherein the leukocyte preparation is produced by pooling multiple preparations produced by the method. ### Claim 16 (depends on 15) The leukocyte preparation of claim 15, wherein the pooling comprises further comprising mixing allogeneic lymphocytes into the leukocyte preparation. ### Claim 17 (depends on 16) The leukocyte preparation of claim 16, wherein the allogeneic lymphocytes are from genetically different individuals of a common species, and wherein the mixing of the allogeneic lymphocytes results in T-cell activation and mediator release for prolonging shelf life of the leukocyte preparation.