# US 11,077,389 B2 · Methods for purifying biological cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,077,389 B2 - Document ID: US11077389B2 - Status: Granted - Kind: B2 - Application number: 16/378,665 - Filed: 2019-04-09 - Published: 2019-08-01 - Granted: 2021-08-03 - Earliest claimed: 2013-02-15 - Assignees: Douglas T. Gjerde - Applicants: Douglas T. Gjerde - Inventors: Douglas T. Gjerde - Web page: https://patentgrep.com/patent/US11077389B2 ## CPC - B01D 15/22: relating to the construction of the column - G01N 1/34: Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)} - B01D 15/14: relating to the introduction of the feed to the apparatus - B01D 15/3809: {of the antigen-antibody type, e.g. protein A, G or L chromatography} - B01D 15/424: {Elution mode} - B01L 3/0275: {Interchangeable or disposable dispensing tips} - G01N 1/405: {by adsorption or absorption} - B01L 2200/0647: Handling flowable solids, e.g. microscopic beads, cells, particles - B01L 2300/069: Absorbents; Gels to retain a fluid - G01N 15/1433: using image recognition - G01N 2015/016: {White blood cells} - G01N 2015/1006: {for cytology} - G01N 2035/0434: {in the form of a syringe or pipette tip} - G01N 2035/1053: {for separating part of the liquid, e.g. filters, extraction phase} - 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) - 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) - B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE ## Abstract This invention relates to devices and methods for purifying biological cells. For example, viable tumor, stem, immune and sperm cells can be purified from a complex biological sample using a pipette tip column. Methods of the invention can aid research, diagnosis and treatment of cancer. ## Claims ### Claim 1 A method for purifying viable cells, comprised of: a) providing at least one fritted column, wherein the fritted column is comprised of a packed bed of medium, wherein the packed bed of medium has a bed volume, wherein the packed bed of medium is retained between two frits, wherein the packed bed of medium is comprised of particles, wherein the particles are comprised of an affinity group, wherein the affinity group is an antibody or a FAB; b) providing a biological sample, wherein the sample is comprised of viable cells, wherein the viable cells are not labeled with antibodies, wherein the biological sample has a sample volume, wherein the sample volume is larger than the bed volume; c) aspirating the biological sample through the fritted column, wherein the sample is aspirated through the packed bed of medium; d) expelling the biological sample by reversing the flow direction, wherein the biological sample is expelled through the packed bed of medium; e) repeating steps (c) and (d) multiple times, wherein the biological sample is not incubated on the fritted column, whereby a portion of the viable cells are captured by the affinity groups on the particles in the packed bed of medium; f) passing a wash solution through the fritted column; and g) eluting the cells by passing an eluent through the fritted column, wherein the eluted cells are intact and viable, wherein the eluted cells maintain their integrity, and wherein the method is at least partially performed by a timed computer-controlled program. ### Claim 2 (depends on 1) The method of claim 1, wherein the fritted column is a pipette tip. ### Claim 3 (depends on 1) The method of claim 1, wherein the sample is comprised of biological fluids or tissues. ### Claim 4 (depends on 1) The method of claim 1, wherein the sample is comprised of bacterial cells, animal cells or plant cells. ### Claim 5 (depends on 1) The method of claim 1, wherein following step (f) the captured cells are labeled on the fritted column. ### Claim 6 (depends on 1) The method of claim 1, wherein the antibodies are covalently attached to the particles with a linker. ### Claim 7 (depends on 1) The method of claim 1, wherein the antibodies are tagged with a drug. ### Claim 8 (depends on 1) The method of claim 1, wherein the particles are comprised of protein A or protein G. ### Claim 9 (depends on 1) The method of claim 1, wherein the method is used for negative selection and viable cells captured by the antibody affinity groups are not the cells of interest. ### Claim 10 (depends on 1) The method of claim 1, wherein the eluted cells are selected from the group consisting of tumor cells, circulating tumor cells, immune cells, sperm cells, white blood cells, cancer cells, E. coli cells, T cells, B cells and stem cells. ### Claim 11 A method for purifying viable cells, comprised of: a) providing at least one fritted column, wherein the fritted column is comprised of a packed bed of medium, wherein the packed bed of medium has a bed volume, wherein the packed bed of medium is retained between two frits, wherein the packed bed of medium is comprised of particles, wherein the particles are comprised of an affinity group, wherein the affinity group is an antibody or a FAB; b) providing a biological sample, wherein the sample is comprised of viable cells, wherein the viable cells are not labeled with antibodies, wherein the biological sample has a sample volume, wherein the sample volume is larger than the bed volume; c) aspirating the biological sample through the fritted column, wherein the sample is aspirated through the packed bed of medium; d) expelling the biological sample by reversing the flow direction, wherein the biological sample is expelled through the packed bed of medium; e) repeating steps (c) and (d) multiple times, wherein the biological sample is not incubated on the fritted column, whereby a portion of the viable cells are captured by the affinity groups on the particles in the packed bed of medium; f) passing a wash solution through the fritted column; and g) eluting the cells by passing an eluent through the fritted column, wherein the eluted cells are intact and viable, wherein the eluted cells maintain their integrity, wherein the method is at least partially performed by a timed computer-controlled program, and wherein the entire method is performed in less than one hour.