Cell-free protein synthesis system
View Patent ↗The technology provided herein relates to novel methods and systems for cell-free protein synthesis, in particular to cell-free protein synthesis systems suitable for protein synthesis before and after freezing for a short- or long-term storage, wherein said system comprises a cryoprotectant.
1. A cell-free protein synthesis system suitable for protein synthesis before and after freezing for a short- or long-term storage, wherein said system comprises a cryoprotectant, wherein said cell-free protein synthesis system is a tobacco BY-2 cell lysate, wherein said cryoprotectant is trehalose or dimethyl sulfoxide (DMSO), wherein a concentration of trehalose is in a range between 1.25 and 2.5% w/v and a concentration of DMSO is in a range between 1 and 4% v/v.
2. The cell-free protein synthesis system according to claim 1 , wherein said cell-free protein synthesis system is frozen.
3. The cell-free protein synthesis system according to claim 1 , wherein said cell-free protein synthesis system is stored at a temperature below −50° C., below −60° C., below −70° C., below −80° C., below −90° C. or below −196° C.
4. A method for cell-free protein synthesis, said method comprising the steps of:
a) freezing the cell-free protein synthesis system according to claim 1 to a temperature below −50° C., below −60° C., below −70° C., below −80° C., below −90° C. or below −196° C.;
b) storing said frozen cell-free protein synthesis system;
c) thawing said cell-free protein synthesis system; and
d) synthesizing a protein in vitro by using said cell-free protein synthesis system and a nucleic acid template.
5. The method according to claim 4 , wherein the cryoprotectant is not removed before synthesizing a protein.
6. The method according to claim 4 , wherein the cell-free protein synthesis system is thawed at room temperature before usage.
7. A method of preserving a cell-free protein synthesis system comprising the steps of:
a) contacting a cell-free protein synthesis system with a cryoprotectant to obtain a cell-free protein synthesis system according to claim 1 ; and
b) freezing said cell-free protein synthesis system in contact with said cryoprotectant.
8. The method according to claim 7 , wherein said cell-free protein synthesis system is frozen by shock freezing.
9. The method according to claim 7 , wherein said cell-free protein synthesis system is frozen to a temperature below −50° C., below −60° C., below −70° C., below −80° C., below −90° C. or below −196° C.
10. The method according to claim 7 , wherein the frozen cell-free protein synthesis system is thawed at room temperature before usage.