Consumable cryopreserved cells transiently overexpressing gene(s) encoding drug transporter protein(s)
The present invention discloses cryopreserved recombinant cells for screening drug candidates that transiently overexpress one or more drug transporter proteins and/or drug metabolizing enzymes. Advantageously, such cells provide a cost-efficient consumable product that streamlines the process of screening whether drug candidates are substrates or inhibitors of drug transporter proteins and/or drug metabolizing enzymes.
1. A recombinant cell comprising one or more transiently overexpressed genes encoding a drug transporter protein, wherein:
the recombinant cell is cryopreserved,
activity of the drug transporter protein is detectable in a population of the recombinant cells prior to cryopreservation, activity of the drug transporter protein is detectable in a population of the recombinant cells following thaw from cryopreservation;
wherein the cryopreserved recombinant cell is transiently transfected with the one or more genes by a method comprising electroporation; and
wherein said one or more genes is OAT3 and wherein said cell is HEK293.
2. The recombinant cell of claim 1 , wherein:
the detectable activity of the drug transporter protein prior to cryopreservation is the activity of the drug transporter protein towards a prototypical substrate for the drug transporter protein, and
the detectable activity in the population of the recombinant cells prior to cryopreservation is at an uptake ratio of from 5 to 25.
3. The recombinant cell of claim 1 , wherein:
the detectable activity of the drug transporter protein following thaw from cryopreservation is the activity of the drug transporter protein towards a prototypical substrate for the drug transporter protein, and
the detectable activity in a population of the recombinant cells following thaw from cryopreservation at an uptake ratio of at least 5.
4. A process of preparing cryopreserved transiently transfected recombinant cells, the process comprising:
transiently transfecting cells with one or more genes encoding a drug transporter protein to provide the transiently transfected recombinant cells, and
cryopreserving the transiently transfected recombinant cells within 48 hours of transient transfection, wherein a population of the transiently transfected recombinant cells transiently overexpress the one or more genes encoding the drug transporter protein at a detectable level prior to cryopreserving the transiently transfected recombinant cells, wherein the transient transfection of the cells comprises electroporation, and wherein said one or more genes is OAT3 and wherein said cell is HEK293.
5. The process of claim 4 , wherein the detectable level prior to cryopreserving is the activity of the drug transporter protein towards a specific prototypical substrate for the drug transporter protein, and wherein the detectable level prior to cryopreserving is an uptake ratio of at least 5.
6. The process of claim 4 , wherein the detectable level prior to cryopreserving is the activity of the drug transporter protein towards a specific prototypical substrate for the drug transporter protein, and wherein the detectable level prior to cryopreserving is an uptake ratio of from 5 to 25.
7. The process of claim 4 , wherein:
a population of the transiently transfected recombinant cells transiently overexpress the one or more genes encoding the drug transporter protein at the detectable level following thaw from cryopreservation,
the detectable level following thaw from cryopreservation is the activity of the drug transporter protein towards a specific prototypical substrate for the drug transporter protein, and
wherein the detectable level following thaw from cryopreservation is an uptake ratio of at least 5.
8. The process of claim 4 , wherein the transiently transfected recombinant cells are cryopreserved at about 24 hours to about 48 hours post transient transfection.