System and method for production of on-demand proteins in a portable unit for point of care delivery

1. A method of producing a composition, wherein the composition comprises a purified on-demand cell-free synthesized protein and a buffer, wherein the composition is free of stabilizers, oxidation/reduction agents, and/or other excipients, said method comprising:

producing the on-demand cell-free synthesized protein in a production module; and

purifying the on-demand cell-free synthesized protein in a purification module,

wherein the production module and the purification module are communicatively connected to one another such that the purification module receives the on-demand cell-free synthesized protein from the production module, and

wherein the purification module comprises a chromatography component for receiving the on-demand cell-free synthesized protein from the production module and for outputting a first purified protein; and a diafiltration component for receiving the first purified protein from the chromatography component and for outputting purified on-demand cell-free synthesized protein, wherein the diafiltration component comprises a product section for receiving the first purified protein from the chromatography component and a buffer section for receiving a buffer solution.

2. The method of claim 1 , wherein the production module comprises a reaction mixture comprising a nucleotide template for production of the on-demand cell-free synthesized protein; monomers for the on-demand cell-free protein to be synthesized; and co-factors, enzymes and reagents that are necessary for synthesis of the on-demand cell-free protein.

3. The method of claim 2 , wherein the enzymes and reagents that are necessary for the synthesis are ribosomes, tRNA, polymerases and transcriptional factors.

4. The method of claim 2 , wherein potency/activity of the on-demand cell-free synthesized protein is at least 55% or more of the initial activity for at least 3 days at temperature from about 0° C. to about 30° C.

5. The method of claim 1 , wherein the purification module and the production module are in a portable bioprocessing system.

6. The method of claim 1 , wherein the composition is refrigerated at an above-freezing temperature from 0 to 6° C. after purification.

7. The method of claim 1 , wherein the production module comprises a bioreactor for cell-free protein expression.

8. The method of claim 2 , wherein the reaction mixture is maintained at a pH between pH 6-9 during production of the on-demand cell-free synthesized protein.

9. The method of claim 2 , wherein the reaction mixture is maintained at a temperature between 20° C. and 40° C. during production of the on-demand cell-free synthesized protein.

10. The method of claim 1 , wherein the diafiltration component further comprises: a first substrate; a second substrate; and a diafiltration membrane positioned between the first and second substrates.

11. The method of claim 10 , wherein the diafiltration component comprises a flow cell.

12. The method of claim 1 , wherein the production module comprises at least one sensor for monitoring at least one of conductivity, temperature, pH, oxygen and CO 2 .

13. The method of claim 1 , wherein the production module and purification module each comprise a heating and cooling element for controlling the temperature of solution inside the production module and purification module.

14. The method of claim 5 , wherein the portable bioprocessing system further comprises a fluid storage and dispensing module for storing solutions used by the production module for protein expression and for storing waste product produced by the purification module.

15. The method of claim 14 , wherein the fluid storage and dispensing module also stores purified on-demand cell-free synthesized protein output by the purification module.

16. The method of claim 5 , wherein the portable bioprocessing system further comprises a processor for controlling and/or monitoring at least one of the production module and the purification module.

17. The method of claim 1 , wherein a therapeutic dose and potency/activity of the on-demand cell-free synthesized protein is determined by expression time in the production module.

18. The method of claim 1 , wherein the composition can be delivered directly to a patient.

19. The method of claim 1 , wherein a therapeutic dose and potency/activity of the on-demand cell-free synthesized protein is determined by the holding time in the production module.

20. The method of claim 1 , wherein the composition is delivered in vivo to a patient in need of such protein within one hour to two weeks after purification of the cell-free synthesized protein.

21. The method of claim 1 , wherein the composition is delivered continuously or as a bolus to a subject in need of the protein comprised therein.