Desiccated biologics and methods of preparing the same
The present invention provides compositions comprising desiccated biologics comprising a cell, protein, virus, nucleic acid, carbohydrate, or lipid, or any combination thereof, along with at least one membrane penetrable sugar, and at least one membrane impenetrable sugar, wherein the moisture content is from 5% to 95%, and to methods of preparing the same, and to methods of treating animals using the same.
1. A method of preserving a cell from a cell line comprising:
contacting the cell with at least one membrane penetrable sugar;
contacting the cell with a neutral dextran with a molecular weight greater than or equal to 50,000 daltons;
optionally, contacting the cell with a fixative agent; and
drying the cell by vacuum desiccation to a final moisture content of from about 5% to about 90%, wherein the % viability of the dried cell upon rehydration is about 30% or greater.
2. The method of claim 1 wherein the membrane penetrable sugar is trehalose.
3. The method of claim 1 wherein the moisture content is from about 15% to about 40%.
4. The method of claim 1 wherein the moisture content is from about 20% to about 25%.
5. The method of claim 1 wherein the fixative agent is glutaraldehyde or paraformaldehyde.
6. The method of claim 1 wherein the cell is dried by vacuum desiccation at temperatures ranging from about 0° C. to about 40° C. for about 1 hours to about 4 hours.
7. The method of claim 6 wherein the cell is dried by vacuum desiccation at temperatures ranging from about 32° C. to about 34° C. for about 3 hours.
8. The method of claim 7 further comprising storing the cell in a vacuum sealed container in the presence or absence of a desiccant.
9. The method of claim 7 further comprising rehydrating the cell.
10. The method of claim 9 wherein the rehydration comprises contacting the cell with water, optionally followed by saline.
11. The method of claim 9 wherein the rehydration comprises contacting the cell with a physiological buffer.
12. The method of claim 11 wherein the physiological buffer is HEPES, phosphate buffered saline, or a Tris buffer.
13. The method of claim 1 wherein the membrane penetrable sugar is trehalose, the membrane impenetrable sugar is dextran, and the moisture content is from about 15% to about 40%.
14. The method of claim 1 wherein the cell line is Chinese hamster ovary cells, 3T3 fibroblasts, or HEK cells.
15. A method of preserving a cell from a cell line comprising:
introducing into the cell at least one membrane penetrable sugar by a method consisting of suspending the cell in a solution comprising the at least one membrane penetrable sugar;
contacting the cell with a neutral dextran with a molecular weight greater than or equal to 50,000 daltons;
optionally, contacting the cell with a fixative agent; and
drying the cell by vacuum desiccation to a final moisture content of from about 5% to about 90%.
16. The method of claim 1 wherein the % viability of the dried cell upon rehydration is about 75% or greater.