# US 2019/0167889 A1 · Automated Method For Leukocyte Collection From Whole Blood > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2019/0167889 A1 - Document ID: US20190167889A1 - Status: Published application - Kind: A1 - Application number: 16/324,178 - Filed: 2017-08-17 - Published: 2019-06-06 - Earliest claimed: 2016-09-06 - Assignees: Fresenius Kabi Deutschland GmbH - Applicants: Fresenius Kabi Deutschland Gmbh - Inventors: Heike Mitschulat; Michael Brinkmann - Web page: https://patentgrep.com/patent/US20190167889A1 ## CPC - A61M 1/3696: {with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation} - A61M 1/0209: {Multiple bag systems for separating or storing blood components} - A61M 1/0272: {Apparatus for treatment of blood or blood constituents prior to or for conservation, e.g. freezing, drying or centrifuging} - A61M 1/382: {Optimisation of blood component yield} - B04B 5/0442: {with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation} - B04B 11/02: Continuous feeding or discharging; Control arrangements therefor - A61M 2202/0407: Lymphocytes - A61M 2202/0437: Blood stem cells - A61M 2202/0439: White blood cells; Leucocytes - A61M 2205/3303: Using a biosensor - A61M 2205/331: used as turbidity change detectors, e.g. for priming-blood or plasma-hemoglubine-interface detection - A61M 2205/3331: Pressure; Flow - B04B 2013/006: {Interface detection or monitoring of separated components} - A61M: DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY (introducing media into or onto the bodies of animals A61D7/00; means for inserting tampons A61F13/26; devices for administering food or medicines orally A61J; containers for collecting, storing or administering blood or medical fluids A61J1/05); DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY (surgery A61B; chemical aspects of surgical articles A61L); DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR {(Electrotherapy, e.g. producing anaesthesia by the use of alternating or intermittent currents A61N1/36021)} - B04B: CENTRIFUGES (high-speed drum mills B02C19/11; domestic spin driers D06F; analysing, measuring or monitoring physical or chemical properties of samples during centrifuging, see the relevant subclasses for these procedures, e.g. G01N) ## Abstract The present invention relates to a method for separating lymphocytes and/or stem cells from whole blood in an automated blood separation system, wherein the quality of the collected lymphocytes and/or stem cells fractions is increased and the cell collection procedure is further automated by use of an optical sensor comprised in a detector device to measure turbidity and colour in the claimed method and in a cell separator, which can be used to perform the claimed method. The method of the invention is particularly useful to collect lymphocytes and/or stem cells fractions from whole blood, wherein the contamination of the collected cell fractions by platelets, red blood cells and granulocytes is reduced. ## Claims ### Claim 1 A method for separating lymphocytes and/or stem cells from whole blood in an automated blood cell separation device, comprising: supplying blood to a separation chamber in a centrifugation unit of the automated blood cell separation device; separating the blood in the separation chamber into red blood cell and plasma fractions, with a buffy-coat including lymphocytes and/or stem cells between the red blood cell and plasma fractions; accumulating the buffy-coat between the red blood cell and plasma fractions in the separation chamber; while portions of the red blood cell and plasma fractions are removed from the separation chamber, with said portion of the plasma fraction removed from the separation chamber flowing through a plasma/buffy-coat tube as part of a fluid stream; causing a spillover in which the buffy-coat and then the red blood cell fraction exits the separation chamber and flows through the plasma/buffy-coat tube after said portion of the plasma fraction as parts of the fluid stream; and transferring the buffy-coat from the plasma/buffy-coat tube into a collection container, wherein a detector device of the automated blood cell separation device monitors at least turbidity and colour of the fluid stream in the plasma/buffy-coat tube, the transfer of the buffy-coat into the collection container is automatically initiated when the detector device determines that the turbidity of the fluid stream in the plasma/buffy-coat tube has risen from a level indicating the presence of the plasma fraction to a level indicating the presence of the buffy-coat and that the colour of the fluid stream in the plasma/buffy-coat tube has changed from a colour indicating the presence of the plasma fraction to a colour indicating the presence of the buffy-coat, and the transfer of the buffy-coat into the collection container is automatically terminated when the detector device determines that a predefined threshold of the colour of the fluid stream in the plasma/buffy-coat tube has been reached, with the predefined threshold of the colour of the fluid stream in the plasma/buffy-coat tube indicating the presence of the red blood cell fraction. ### Claim 2 (depends on 1) The method of claim 1, wherein the detector device comprises an optical sensor. ### Claim 3 (depends on 2) The method according to claim 2, wherein the optical sensor comprises a red LED for turbidity measurement and a green LED for measuring absorption of green light by the red blood cell fraction in the plasma/buffy-coat tube. ### Claim 4 (depends on 3) The method according to claim 3, further comprising monitoring at least a portion of the light emitted by the LEDs to enable compensation for aging effects and temperature drift in the LEDs. ### Claim 5 (depends on 1) The method according to claim 1, wherein the detector device is configured to, upon detecting the increase of the turbidity and the change of the colour of the fluid stream in the plasma/buffy-coat tube, transmit a signal to a processor device, and the processor device is configured to, upon receiving the signal from the detector device, initiate the transfer and collection of the buffy-coat by switching a position of a spillover clamp associated with the plasma/buffy-coat tube so as to direct the fluid stream in the plasma/buffy-coat tube into the collection container. ### Claim 6 (depends on 1) The method according to claim 1, wherein the detector device is configured to, upon determining that the colour of the fluid stream in the plasma/buffy-coat tube has reached the predefined threshold, transmit a signal to a processor device, and the processor device is configured to, upon receiving the signal from the detector device, terminate the transfer and collection of the buffy-coat by changing a position of a spillover clamp associated with the plasma/buffy-coat tube so as to direct the fluid stream in the plasma/buffy-coat tube away from the collection container. ### Claim 7 (depends on 1) The method according to claim 1, wherein the predefined threshold of the colour of the fluid stream in the plasma/buffy-coat tube corresponds to a red colour of a fluid having a hematocrit value of up to 5%. ### Claim 8 (depends on 1) The method according to claim 1, wherein the collection of the buffy-coat is terminated after detection of the predefined threshold of the colour of the fluid stream in the plasma/buffy-coat tube and after an additional default volume of the buffy-coat is allowed to pass a spillover clamp associated with the plasma/buffy-coat tube. ### Claim 9 (depends on 8) The method according to claim 8, wherein said accumulating the buffy-coat in the separation chamber, said causing a spillover, and said transferring the buffy-coat are repeated in a plurality of cycles, in at least one of the cycles, after detection of the predefined threshold of the colour of the fluid stream in the plasma/buffy-coat tube, said additional default volume of the buffy-coat is allowed to pass the spillover clamp before the collection of the buffy-coat for said at least one of the cycles is terminated, and in at least one subsequent cycle, after detection of the predefined threshold of the colour of the fluid stream in the plasma/buffy-coat tube, a reduced additional default volume of the buffy-coat is allowed to pass the spillover clamp before the collection of the buffy-coat for said at least one subsequent cycle is terminated. ### Claim 10 (depends on 1) The method according to claim 1, wherein the automated blood cell separation device is directly connected to a donor/patient and the blood, which is to be separated, is obtained via a needle from the donor/patient and supplied directly to the automated blood cell separation device, and the plasma and red blood cell fractions are reinfused into the donor/patient via a second needle. ### Claim 11 (depends on 1) The method according to claim 1, wherein the automated blood cell separation device is not connected to a patient, said method is performed using blood samples that were already obtained from the patient, and the plasma and red blood cell fractions are not reinfused into the patient but are collected in collection containers. ### Claim 12 (depends on 1) The method according to claim 1, wherein a product collected in the collection container substantially contains lymphocytes and/or stem cells. ### Claim 13 (depends on 1) The method according to claim 1, wherein a product collected in the collection container has a reduced count of platelets, granulocytes and red blood cells or is substantially free of platelets, granulocytes and red blood cells. ### Claim 14 (depends on 13) The method according to claim 13, wherein a product collected in the collection container k has a hematocrit value, which is below 10% when stem cells are collected as the product, and below 5% when lymphocytes are collected as the product. ### Claim 15 A blood cell separation device for separating lymphocytes and/or stem cells from whole blood in an automated procedure, comprising: a pumping section for pumping a fluid through a tubing set including a plasma/buffy-coat tube installed on the blood cell separation device; a centrifuge compartment, which comprises a separation chamber for the automated separation of buffy-coat from whole blood; a detector device for determining the presence of target cells, such as lymphocytes and/or stem cells, in a fluid stream in the plasma/buffy-coat tube, and for determining the presence of red blood cells in the fluid stream in the plasma/buffy-coat tube; an automatic clamp for directing the fluid stream in the plasma/buffy-coat tube in a direction away from a collection container and for directing the fluid stream in the plasma/buffy-coat tube into the collection container; and a processor device for controlling the automatic clamp, the processor device being constituted to switch the position of the automatic clamp to direct the fluid stream in the plasma/buffy-coat tube into the collection container when the detector device determines the target cells to be present in the plasma/buffy-coat tube (124), and the processor device being further constituted to switch the position of the automatic clamp to direct the fluid stream in the plasma/buffy-coat tube away from the collection container when the detector device determines the red blood cells to be present in the plasma/buffy-coat tube, wherein the detector device monitors at least turbidity and colour of the fluid stream in the plasma/buffy-coat tube and the transfer of the target cells into the collection container is initiated when the detector device determines that the turbidity of the fluid stream in the plasma/buffy-coat tube has risen from a level indicating the absence of the target cells to a level indicating the presence of the target cells and that the colour of the fluid stream in the plasma/buffy-coat tube has changed from a colour indicating the absence of the target cells to a colour indicating the presence of the target cells, and the transfer of the target cells into the collection container is terminated when the detector device determines that a predefined threshold of the colour of the fluid stream in the plasma/buffy-coat tube has been reached, with the predefined threshold of the colour of the fluid stream in the plasma/buffy-coat tube indicating the presence of red blood cells. ### Claim 16 (depends on 1) The method according to claim 1, wherein said causing a spillover comprises increasing a rate at which the plasma fraction is removed from the separation chamber and reducing a rate at which blood is supplied to the separation chamber. ### Claim 17 (depends on 1) The method according to claim 1, wherein the predefined threshold of the colour of the fluid stream in the plasma/buffy-coat tube corresponds to a red colour of a fluid having a hematocrit value of up to 25%. ### Claim 18 (depends on 9) The method of claim 9, wherein said additional default volume of the buffy-coat is allowed to pass the spillover clamp before the collection of the buffy-coat for the cycle is terminated for the first half of the cycles to be executed, and said reduced additional default volume of the buffy-coat is allowed to pass the spillover clamp before the collection of the buffy-coat for the cycle is terminated for the second half of the cycles to be executed. ### Claim 19 (depends on 9) The method of claim 9, wherein the reduced additional volume of the buffy-coat is approximately half of the additional default volume of the buffy-coat. ### Claim 20 (depends on 14) The method of claim 14, wherein the product collected in the collection container has a hematocrit value which is below 8% when stem cells are collected as a product and below 2% when lymphocytes are collected as the product.