Protein expression yield enhancement in cell-free protein synthesis systems by addition of antifoam agents
AssigneeThe Board of Trustee of the Leland Stanford Junior University
InventorsAlexei M. Voloshin, James Robert Swartz
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Abstract
Compositions and methods are provided for the in vitro synthesis of biological molecules in reaction mixtures comprising anti-foam agents. The reaction mix comprising antifoam agent may be a scaled up reaction, e.g. in reaction volume greater than at least about 15 μl. Reactions may be performed in various reactors, as known in the art, which include stirred reactors, bubble-column reactors; and the like.
Claims13
A method for translation of mRNA to produce polypeptides, the method comprising:
- synthesizing said polypeptides in a cell-free reaction mixture of greater than about 15 μl volume, comprising an antifoam agent at a concentration of at least 0.00007%, and not more than 0.007% by weight, wherein the antifoam agent is other than a detergent.
A method for translation of mRNA to produce polypeptides, the method comprising:
- synthesizing said polypeptides in a cell free reaction mixture of greater than about 15 μl volume, comprising:
- a cell extract;
- a template for production of the mRNA and/or polypeptides;
- monomers for the mRNA and/or polypeptides to be synthesized;
- and co-factors, enzymes and reagents that are necessary for the synthesis;
- and an anti-foam agent at a concentration of at least 0.00007%, and not more than 0.007% by weight, wherein the antifoam agent is other than a detergent.
A reaction mixture for cell-free translation of mRNA to produce polypeptides, comprising:
- a cell extract;
- mRNA;
- monomers for the polypeptide to be synthesized;
- and co-factors, enzymes and reagents that are necessary for the synthesis;
- and an anti-foam agent other than a detergent at a concentration of at least 0.00007%, and not more than 0.007% by weight.
Dependencies are inferred from the claim text.