# US 12,180,444 B2 · Containers and methods for cell transduction > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,180,444 B2 - Document ID: US12180444B2 - Status: Granted - Kind: B2 - Application number: 17/136,400 - Filed: 2020-12-29 - Published: 2021-07-01 - Granted: 2024-12-31 - Earliest claimed: 2019-12-30 - Assignees: Saint-Gobain Performance Plastics Corporation - Applicants: Saint-Gobain Performance Plastics Corporation - Inventors: Katie Campbell; Natalie Fekete; Rachel Z. Pytel - Web page: https://patentgrep.com/patent/US12180444B2 ## CPC - C12M 23/14: {Bags} - C12M 23/20: {Material Coatings (immunocoatings C12M25/00)} - C12M 25/00: {Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings} - C12M 35/00: {Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion (machines for extracting juice from animal or plant tissue by electroplasmolysis A23N1/006, processes employing electric or wave energy B01J19/08; treatment of microorganisms or enzymes with electrical or wave energy C12N13/00; methods for cell fusion C12N15/02; introduction of foreign genetic material C12N15/87)} - C12N 15/86: Viral vectors - C12N 15/87: Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation - 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} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract This disclosure relates generally to containers (such as bags) having surfaces comprising one or more aptamer sequences. More particularly, the present disclosure relates to containers such as bags comprising a fluoropolymer attached to aptamer sequences having binding affinity for one or more biological agents, and to transductions methods using such containers. In one aspect, the disclosure provides a container (e.g., a bag) having an outer surface and an inner surface, the inner surface comprising a fluoropolymer; attached to the fluoropolymer, a plurality of functional groups; and attached to each of at least a portion of the functional groups, a first aptamer sequence having a binding affinity for a viral vector. ## Claims ### Claim 1 A method for introducing DNA to a target cell via transduction with a viral vector that is a lentivirus vector or a retrovirus vector, the method comprising providing a container in the form of a cell culture bag having an outer surface and an inner surface, the inner surface comprising a fluorinated ethylene propylene fluoropolymer; attached to the fluoropolymer, a plurality of functional groups; attached to the fluoropolymer by covalent bonding to at least a first portion of the functional groups, a first nucleotide aptamer sequence having a binding affinity for the viral vector; and attached to the fluoropolymer by covalent bonding to at least a second portion of the functional groups, a second nucleotide aptamer sequence having a binding affinity for a VLA-4 or VLA-5 cell receptor of the target cell; and incubating the viral vector and the target cell in the container, such that the viral vector is bound to the first nucleotide aptamer sequence and the target cell is bound to the second nucleotide aptamer sequence. ### Claim 2 (depends on 1) The method of claim 1, wherein the first nucleotide aptamer sequence and/or the second nucleotide aptamer sequence is provided by an oligonucleotide comprising the first nucleotide aptamer sequence and/or the second nucleotide aptamer sequence directly attached to a functional group of the surface. ### Claim 3 (depends on 1) The method of claim 1, wherein the first nucleotide aptamer sequence and/or the second nucleotide aptamer sequence is provided by a polynucleotide comprising two or more repeats of the first nucleotide aptamer sequence and/or the second nucleotide aptamer sequence directly attached to a functional group of the surface. ### Claim 4 (depends on 3) The method of claim 3, wherein the polynucleotide comprises at least one repeat of each of the first aptamer sequence and the second aptamer sequence. ### Claim 5 (depends on 3) The method of claim 3, wherein the polynucleotide comprises at least 10 repeats of the first aptamer sequence or the second aptamer sequence. ### Claim 6 (depends on 1) The method of claim 1, wherein the first nucleotide aptamer sequence and/or the second nucleotide aptamer sequence is provided by at least one oligonucleotide comprising the first nucleotide aptamer sequence and at least one oligonucleotide comprising the second nucleotide aptamer sequence, each attached to a functional group of the surface through a linker, wherein at least 10 oligonucleotides are attached to the surface. ### Claim 7 (depends on 1) The method of claim 1, wherein the functional groups are the product of etching of the fluoropolymer; the product of activation of the fluoropolymer in the presence of a reactive species; or the product of chemically treating an activated fluoropolymer. ### Claim 8 (depends on 1) The method of claim 1, wherein the functional groups include one or more of carboxy groups, hydroxyl groups, aldehyde groups, amine groups, imine groups, amide groups and ester groups. ### Claim 9 (depends on 1) The method of claim 1, wherein the functional groups include amine groups. ### Claim 10 (depends on 1) The method of claim 1, wherein the viral vector comprises a lentivirus vector. ### Claim 11 (depends on 1) The method of claim 1, comprising including a suspension of the viral vector in a first aqueous medium to the container; incubating the container comprising the viral vector for a first period of time; adding the target cell to the container; incubating the container comprising the target cell and the viral vector for a second period of time; and then collecting transduced cells from the container. ### Claim 12 (depends on 11) The method of claim 11, further comprising, after incubating the container for the first period of time removing at least a portion of the first aqueous media; then adding a wash media to the container; then removing at least a portion of the wash media from the container; and then adding the target cell to the container. ### Claim 13 (depends on 11) The transduction method of claim 11, wherein collecting transduced cells from the container comprises adding a cell dissociation media to the container; and then removing a suspension of transduced cells from the container. ### Claim 14 (depends on 1) The method of claim 1, comprising including a suspension of the viral vector and the target cell in a first aqueous media to the container; incubating the container comprising the target cell and the viral vector for a first period of time; and then collecting transduced cells from the container. ### Claim 15 (depends on 1) The method of claim 1, wherein the viral vector comprises a retrovirus vector. ### Claim 16 (depends on 1) The method of claim 1, wherein the functional groups are the product of activation of the fluoropolymer by plasma activation or corona activation in the presence of a reactive hydrocarbon species.