# US 11,994,528 B2 · Systems, methods, and devices for automated nucleic acid and protein isolation > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,994,528 B2 - Document ID: US11994528B2 - Status: Granted - Kind: B2 - Application number: 17/122,324 - Filed: 2020-12-15 - Published: 2021-06-24 - Granted: 2024-05-28 - Earliest claimed: 2019-12-18 - Assignees: Life Technologies Corporation - Applicants: Life Technologies Corporation - Inventors: Alexandre Vlassov; Timothy Barta; Justin Inslee; Ethan Gentert; Robert Smith; Sara Thiele; Carolyn Adams; Kraig Kruger; Robert Schneider; Wade Lukianow; Jay Greco; Jason Jones; David Cerrone; Sumonto Ghosh; Brian Vajda; Andrew Basta; Allan Evans; Christopher Crowley; Robert Setterquist; Anne Cox; Mark Mayernick - Web page: https://patentgrep.com/patent/US11994528B2 ## CPC - C12N 15/1003: {Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor} - G01N 35/1016: {Control of the volume dispensed or introduced} - B01D 61/58: Multistep processes - B01D 69/02: characterised by their properties - C12M 47/06: {Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material (lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003)} - C12M 47/10: {Separation or concentration of fermentation products (bioreactors combined with means for distillation or extraction of liquid fuel C12M43/02)} - C12M 47/12: {Purification (C12M47/04 takes precedence)} - G01N 1/405: {by adsorption or absorption} - G01N 1/4077: {by other techniques involving separation of suspended solids} - G01N 21/59: Transmissivity (G01N21/25 takes precedence) - G01N 35/00: Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor - G01N 35/00584: {Control arrangements for automatic analysers} - G01N 35/1081: {characterised by the means for relatively moving the transfer device and the containers in an horizontal plane (G01N35/1011 takes precedence)} - G01N 35/1095: {for supplying the samples to flow-through analysers (for a specific analyser see relevant groups, e.g. under G01N15/00, G01N21/00, G01N27/00, G01N30/00, H01J49/00)} - B01D 2311/2642: Aggregation, sedimentation, flocculation, precipitation or coagulation - B01D 2313/44: Cartridge types - B01D 2313/50: Specific extra tanks - B01D 2313/58: Parts of membrane modules specifically adapted for single use - B01D 2319/025: Permeate series - B01D 2325/42: Ion-exchange membranes - G01N 2001/4088: {filtration} - G01N 2035/00316: {Detecting door closure} - G01N 2035/0436: {with pre-packaged reagents, i.e. test-packs} - G01N 2035/0491: {Position sensing, encoding; closed-loop control} - G01N 2035/1025: {Fluid level sensing} - G01N 2035/1053: {for separating part of the liquid, e.g. filters, extraction phase} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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) - 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) - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} ## Abstract Purifying target biomolecules, such as nucleic acids or proteins, from a biological source is a time intensive process and is typically performed by a skilled technician or scientist owing to the highly technical nature of the work. Systems, devices, and methods disclosed herein enable the automated bioprocessing and purification of target biomolecules from a biological source. For example, an instrument and disposable cartridge are provided for automatedly isolating and purifying nucleic acids (such as plasmid DNA from a bacterial culture) or for isolating protein from any biological sample. Such an exemplary instrument and cartridge can work in concert to timely release, mix, and move the target biomolecule and various reagents and buffers through a target biomolecule purification process, resulting in a purified target biomolecule with less manual oversight than traditional approaches. ## Claims ### Claim 1 An instrument for automated purification of a target biomolecule from a biological sample, comprising: a purification cartridge comprising an apparatus that includes: an input reservoir for receiving the biological sample; a first bioprocessing assembly in fluid communication with the input reservoir and a lysis buffer reservoir, the first bioprocessing assembly configured to generate a lysate comprising the target biomolecule; a second bioprocessing assembly in fluid communication with the first bioprocessing assembly and a first elution buffer reservoir, the second bioprocessing assembly including a target-biomolecule-binding filter configured to retain the target biomolecule; and a receptacle in fluid communication with the second bioprocessing assembly, the receptacle configured to receive an output container for receiving the target biomolecule from the second bioprocessing assembly, the purification cartridge further comprising: a first external layer comprising a first side and a second side, and a second external layer disposed opposite the first side of the first external layer; a casing having an internal compartment configured in size and shape for receiving the purification cartridge; wherein the internal compartment comprises a position sensor for determining that the purification cartridge has been fully inserted within the internal compartment; optionally, a selectively closable access door providing access to the internal compartment; a pump assembly disposed within the internal compartment and configured to provide pumping action through peristaltic motion; and a clamp mechanism disposed within the internal compartment and configured to move between an open position in which the internal compartment is accessible and a closed position in which the clamp mechanism is clamped against an inserted purification cartridge to enable processing of the biological sample, wherein when in the closed position, the clamp mechanism operates to fluidically seal the purification cartridge by exerting the first external layer and the second external layer towards one another. ### Claim 2 (depends on 1) The instrument of claim 1, further comprising a user interface for displaying instrument information and for receiving user input, the user interface configured to receive user input related to one or more of a volume of biological sample, a selected purification protocol, operating instructions for the selectively closable door, a desired concentration of the target biomolecule, or a final volume of eluent comprising the target biomolecule. ### Claim 3 (depends on 1) The instrument of claim 1, further comprising one or more access door sensors configured to determine whether the access door is in an open or closed position, wherein the one or more access door sensors optionally comprise one or more Hall effect sensors and one or more corresponding magnets. ### Claim 4 (depends on 1) The instrument of claim 1, wherein the pump assembly comprises a camshaft coupled to a motor via a power transmission assembly, a plurality of cam elements being attached to the camshaft and extending transversely therefrom toward the purification cartridge when inserted, the cam elements optionally including beveled tips for engaging the purification cartridge when inserted. ### Claim 5 (depends on 1) The instrument of claim 1, wherein the biological sample is a bacterial culture, a cell culture, a prokaryotic cell culture, a eukaryotic cell culture, a clinical sample, an environmental sample, a food sample, or a beverage sample. ### Claim 6 (depends on 1) The instrument of claim 1, wherein the apparatus further comprises an optical density detector window disposed between the input reservoir and the first bioprocessing assembly, the optical density detector window being configured to allow detection of an optical density of the biological sample. ### Claim 7 (depends on 1) The instrument of claim 1, wherein the first bioprocessing assembly comprises a clarification filter. ### Claim 8 (depends on 7) The instrument of claim 7, wherein the first bioprocessing assembly comprises a cell capture filter, wherein the cell capture filter is disposed upstream of the clarification filter. ### Claim 9 (depends on 1) The instrument of claim 1, wherein the target-biomolecule-binding filter of the second bioprocessing assembly comprises a silica-based filter or a column of beads having affinity for the target biomolecule. ### Claim 10 (depends on 9) The instrument of claim 9, wherein the second bioprocessing assembly further comprises a purification reagent reservoir in fluid communication with the target-biomolecule-binding filter or a protein binding filter. ### Claim 11 (depends on 1) The instrument of claim 1, wherein the second bioprocessing assembly comprises a mixing chamber disposed between the first bioprocessing assembly and the target-biomolecule-binding filter. ### Claim 12 (depends on 11) The instrument of claim 11, wherein the mixing chamber is in fluid communication with an endotoxin removal buffer reservoir. ### Claim 13 (depends on 1) The instrument of claim 1, wherein the output container is selectively detachable from the apparatus. ### Claim 14 (depends on 1) The instrument of claim 1, wherein the input reservoir is sized and shaped to receive up to 2 L of the biological sample. ### Claim 15 (depends on 1) The instrument of claim 1, wherein the target biomolecule is a target nucleic acid, wherein: the first bioprocessing assembly is configured to receive the biological sample, the first bioprocessing assembly comprising a waste-separation filter and a plurality of reservoirs fluidically coupled to the waste-separation filter; the second bioprocessing assembly comprising an anion exchange membrane, a washing solution reservoir fluidically coupled to the anion exchange membrane and the first elution buffer reservoir being fluidically coupled to the anion exchange membrane; and the purification cartridge comprising a third bioprocessing assembly comprising a precipitation filter and a second elution buffer reservoir fluidically coupled to the precipitation filter, wherein the first bioprocessing assembly additionally comprises a cell capture filter, and wherein a first reservoir of the plurality of reservoirs comprises a resuspension buffer reservoir, and wherein the input reservoir is fluidically coupled to a first side of the cell capture filter and the resuspension buffer reservoir is fluidically coupled to a second side of the cell capture filter. ### Claim 16 (depends on 1) The instrument of claim 1, wherein the instrument includes a selectively closable access door providing access to the internal compartment and wherein when in the closed position, the clamp mechanism operates to fluidically seal the purification cartridge and further comprising a controller, wherein the controller routes power to the clamp mechanism only upon determining that the access door is closed. ### Claim 17 (depends on 1) The instrument of claim 1, wherein the input reservoir is sized and shaped to receive between about 5 mL and 5 L of the biological sample. ### Claim 18 (depends on 1) The instrument of claim 1, wherein the input reservoir is sized and shaped to receive between about 100 mL and 1 L of the biological sample.