Emulsion compositions
View Patent ↗An emulsion is useful in allowing a wide variety of gene products to be expressed via eukaryotic in vitro expression. The emulsion comprises a silicone based surfactant, a hydrophobic phase and a hydrophilic phase; wherein the hydrophilic phase comprises a plurality of compartments containing a functional in vitro eukaryotic expression system.
1. A method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising the steps of: (a) compartmentalising the genetic elements into microcapsules formed from an emulsion; (b) expressing the genetic elements to produce their respective gene products within the microcapsules; and (c) sorting the genetic elements which produce gene product(s) having the desired activity, wherein said emulsion comprises a surfactant, a hydrophobic phase, a hydrophilic phase, and a plurality of microcapsules containing a functional in vitro eukaryotic expression system, and wherein the surfactant is a chemically inert silicone-based surfactant.
2. The method of claim 1 , wherein the chemically inert silicone-based surfactant is a silicone copolymer.
3. The method of claim 1 , wherein the chemically inert silicone-based surfactant is polysiloxane-polycetyl-polyethylene glycol copolymer.
4. The method of claim 1 , wherein the surfactant is provided at a v/v concentration in the hydrophobic phase of 4%.
5. The method of claim 1 , wherein the emulsion comprises an in vitro eukaryotic expression system.
6. The method of claim 1 , wherein the eukaryotic expression system comprises all or part of a mammalian cell lysate.
7. The method of claim 5 , wherein the eukaryotic expression system comprises all or part of a mammalian cell lysate.
8. The method of claim 6 , wherein the mammalian cell lysate is a reticulocyte lysate.
9. The method of claim 6 , wherein the mammalian cell lysate is a rabbit reticulocyte lysate.
10. The method of claim 1 , wherein the emulsion is stable during incubation at 30° C. for at least one hour.
11. The method of claim 1 , wherein the emulsion is stable during incubation at 30° C. for at least one hour.
12. The method of claim 1 , wherein the hydrophobic phase comprises a mineral oil.
13. The method of claim 1 , said microcapsules further comprising genetic elements that are capable of being expressed by said expression system to form a gene product.
14. The method of claim 13 , wherein the expression of a gene product within the emulsion is at a level of at least 1% of that of the gene product achievable by the expression system prior to formation of the emulsion.
15. The method of claim 14 , wherein the expression of the gene product within the emulsion is at a level of at least 10%, at least 20%, (Original) or at least 30% of that of the gene product achievable with the expression system prior to formation of the emulsion.
16. The method of claim 1 , wherein said emulsion is formed comprising agitating together in the presence of a chemically inert silicone-based surfactant a hydrophobic composition and a hydrophilic composition comprising a eukaryotic expression system.
17. The method according to claim 16 , wherein the surfactant is present in the hydrophobic composition.