# US 2024/0201051 A1 · Method for collecting cell types > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2024/0201051 A1 - Document ID: US20240201051A1 - Status: Published application - Kind: A1 - Application number: 18/540,020 - Filed: 2023-12-14 - Published: 2024-06-20 - Earliest claimed: 2022-12-16 - Assignees: Flaskworks, LLC - Applicants: Flaskworks, LLC - Inventors: Andrew Kozbial; Lekhana Bhandary - Web page: https://patentgrep.com/patent/US20240201051A1 ## CPC - B04B 5/0442: {with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation} - G01N 1/18: with provision for splitting samples into portions (G01N1/12, G01N1/14 take precedence; fraction-collection apparatus for chromatography B01D15/08) - A61M 1/3693: {using separation based on different densities of components, e.g. centrifuging} - A61M 1/3696: {with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation} - B01D 21/262: {by using a centrifuge} - B01D 2221/10: Separation devices for use in medical, pharmaceutical or laboratory applications, e.g. separating amalgam from dental treatment residues - B04B 2005/0471: {with additional elutriation separation of different particles} - 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) - 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) - 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)} - B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28) ## Abstract The present invention provides methods for collecting desired cells from a sample using counterflow centrifugal elutriation. Methods of the invention provide for a continuous elutriation workflow with optimized conditions that improves cell purity and retention during the elutriation process. Specifically, methods of the invention use a continuous counterflow centrifugal elutriation process that does not require harvesting cells or removing cells from the separation chamber between elutriation cycles. Methods of the invention increase the purity of desired cell populations, cell number yield, cell recovery, and cell viability to improve downstream processing. Methods of the invention also reduce the volume of buffers and reagents necessary for processing and speeds up the elutriation process for a cost-effective and more efficient cell manufacturing workflow. ## Claims ### Claim 1 A method for collecting cells, the method comprising: introducing a first cell load into a rotatable separation chamber of a reverse flow centrifugation device, wherein the first cell load comprises a first set of cells of interest and a first set of one or more other cell types; performing a first elutriation cycle on the first cell load to form a fluidized bed of the first set of cells of interest within the separation chamber while the first set of one or more other cell types are separated apart from the first set of cells of interest; introducing into the separation chamber a second cell load while the first set of cells of interest remains within the separation chamber, wherein the second cell load comprises a second set of cells of interest and a second set of one or more other cell types; and performing a second elutriation cycle on the second cell load to collect the second set of cells of interest on the fluidized bed of the first set of cells of interest in the separation chamber while the second set of one or more other cell types are separated apart from the first and second sets of cells of interest. ### Claim 2 (depends on 1) The method of claim 1, wherein the first elutriation cycle and the second elutriation cycle are performed as a continuous elutriation process. ### Claim 3 (depends on 1) The method of claim 1, further comprising tuning one or more parameters of at least one of the first elutriation cycle or second elutriation cycle to optimize the formation of the fluidized bed of cells of interest and/or the collection of cells of interest on the fluidized bed within the separation chamber. ### Claim 4 (depends on 3) The method of claim 3, wherein the one or more parameters comprise at least one of a rotation speed of the separation chamber, a cell load volume, a cell load flow rate, and a time for fluidized bed formation. ### Claim 5 (depends on 4) The method of claim 4, wherein one or more parameters of the first elutriation cycle are tuned to optimize the formation of the fluidized bed of the first set of cells of interest within the separation chamber. ### Claim 6 (depends on 4) The method of claim 4, wherein one or more parameters of a second elutriation cycle are tuned to optimize the collection of the second set of cells of interest on the fluidized bed of the first set of cells of interest in the separation chamber. ### Claim 7 (depends on 6) The method of claim 6, wherein one or more parameters of the second elutriation cycle are tuned so as to maintain the fluidized bed of the first set of cells of interest within a first portion of the separation chamber while further collecting the second set of cells of interest on the fluidized bed. ### Claim 8 (depends on 7) The method of claim 7, wherein a rotation speed of the separation chamber is adjusted from a first rotation speed associated with performing the first elutriation cycle to a second rotation speed associated with performing the second elutriation cycle that is different than the first rotation speed. ### Claim 9 (depends on 8) The method of claim 8, wherein the second rotation speed results in movement of the fluidized bed of the first set of cells of interest to the first portion of the separation chamber such that a space in the separation chamber is available for collecting the second set of cells of interest on the fluidized bed. ### Claim 10 (depends on 1) The method of claim 1, further comprising performing a final elutriation cycle. ### Claim 11 (depends on 10) The method of claim 10, wherein the final elutriation cycle is performed without prior introduction of an associated cell load into the separation chamber. ### Claim 12 (depends on 1) The method of claim 1, wherein the first cell load and the second cell load are introduced into the separation chamber within an associated fluid. ### Claim 13 (depends on 12) The method of claim 12, wherein each of the first cell load and the second cell load comprise cells suspended within a buffer solution. ### Claim 14 (depends on 13) The method of claim 13, wherein the cells are obtained from an apheresis process. ### Claim 15 (depends on 1) The method of claim 1, further comprising washing an aggregate of cells of interest collected within the separation chamber upon completion of all elutriation cycles. ### Claim 16 (depends on 1) The method of claim 1, further comprising one or more additional elutriation cycles. ### Claim 17 (depends on 1) The method of claim 1, wherein the cells of interest are selected from the group consisting of red blood cells, neutrophils, eosinophils, basophils, monocytes, lymphocytes, and macrophages. ### Claim 18 (depends on 17) The method of claim 17, wherein the cells of interest are monocytes.