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-nutrititive 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 handling a biological drug composition with living cells, the method comprising:
formulating living CD4+ T-cells cells by reactivating the living CD4+ T-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;
storing the reactivated living cells with the agents in the state of activation in a non-nutritive buffer at a storage temperature below about 20° C. for 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 storage in the non-nutritive buffer for greater than about 24 hours in the state of activation in the non-nutritive buffer.
2. The method of claim 1 wherein the storage temperature is in a range between about 0° C. and about 10° C. and the reactivated living cells are stored for greater than 36 hours.
3. The method of claim 1 wherein the concentration of the cells in the non-nutritive buffer is about 10 6 cells/ml or greater.
4. 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.
5. The method of claim 1 wherein the living cells in the composition express CD40L after storage in an amount of at least about 80% relative to the expression of CD40L at the time of formulation.
6. The method of claim 4 wherein the inflammatory cytokine secreted is IFN-γ and the level of secretion is in an amount of at least about 80% compared to the levels at the time of formulation.
7. The method of claim 6 wherein the secretion of IFN-γ after storage is recovered after incubation at 37° C. for at least 24 hours.
8. The method of claim 1 wherein the living cells in the non-nutritive buffer maintain at least about 50% of the activity of the functional characteristic for at least about 72 hours.
9. The method of claim 1 wherein the living cells are placed in a flexible container or syringe, wherein the flexible container or syringe is packaged in a temperature controlled device that maintains the living cells at the storage temperature.
10. The method of claim 9 further comprising shipping and distributing the package in the temperature controlled device to the point of care.
11. The method of claim 1 wherein the composition has at least about 80% viable cells after storage.
12. The method of claim 1 wherein the living cells in the non-nutritive buffer maintain at least about 50% of the activity of the functional characteristic for at least about 24 hours.
13. The method of claim 1 wherein the living cells in the non-nutritive buffer maintain at least about 50% of the activity of the functional characteristic for at least about 72 hours.
14. The method of claim 1 wherein the non-nutritive buffer is a formulation buffer comprising Plasmalyte A and 1% human serum albumin.