Methods for handling biological drugs containing living cells
The present invention includes methods for handling live cell compositions in non-nutritive buffer. The cells in the compositions maintain their identity and functional characteristics after being stored in non-nutritive media up to about 72 hours. The storage method enables the cells to be manufactured at a processing facility and shipped to a point of care site. The invention also includes compositions that have been stored in non-nutritive buffer at storage temperatures while maintaining the functional characteristics.
1. A method of providing living cell compositions to a point of care facility comprising:
formulating living CD4+ T-cells cells in a non-nutritive buffer by reactivating the living CD4+ T-cells cells to a state of activation by cross-linking the CD3 and CD28 cell surface moieties by anti-CD3 and anti-CD28 monoclonal antibodies, wherein the living CD4+ T-cells have been activated by cross-linking CD3 and CD28 cell surface moieties by anti-CD3 and anti-CD28 monoclonal antibodies and wherein the living cells having been more than minimally manipulated, cryopreserved and thawed prior to reactivation at a processing facility;
transporting the reactivated cells to a point of care facility in a package equipped to maintain a storage temperature at 20° C. or below; and
storing the reactivated living cells with the agents in the state of activation in a non-nutritive buffer at a storage temperature of 20° C. or below, wherein the transportation and storage time is greater than 24 hours, wherein the reactivated living cells with the agents maintain a cell marker indicative of the living cells identity, maintain at least one functional characteristic exhibited by the living cells and are useful for immunotherapy after transportation and storage in the non-nutritive buffer for greater than 24 hours in the state of activation in the non-nutritive buffer.
2. The method of claim 1 further comprising placing the formulated cells in a flexible container or syringe prior to transporting.
3. The method of claim 1 wherein the live cells in the composition express CD40L after transportation and removal from storage temperature in an amount of at least about 80% relative to the expression of CD40L at the time of formulation.
4. The method of claim 1 wherein the cells in the composition secrete IFN-γ after transportation and removal from storage temperature in an amount of at least about 80% compared to the levels at the time of formulation.
5. The method of claim 4 wherein the secretion of IFN-γ after transportation and storage is recovered after incubation at 37° C. for at least 24 hours.
6. The method of claim 1 wherein the live cells in the non-nutritive buffer are at the storage temperature for at least about 72 hours.
7. The method of claim 1 wherein the storage temperature is in a range between about 0° C. and 10° C.
8. The method of claim 7 further comprises returning the cell composition to about room temperature prior to administration to a patient.
9. The method of claim 1 wherein the concentration of the cells in the non-nutritive buffer is about 10 6 cells/ml or greater.
10. The method of claim 1 wherein the functional characteristic is expression of CD40L, FasL, perforin and granzyme B, expression of costimulatory molecules, expression of adhesion molecules, secretion of cytokines, chemokines or combinations thereof.
11. The method of claim 1 wherein the composition has at least about 80% cell viability after transportation and removal from the storage temperature.
12. The method of claim 1 wherein the live cells in the non-nutritive buffer are at the storage temperature for at least about 48 hours.
13. The method of claim 1 wherein the living cells are placed in a flexible container or syringe prior to packaging.