Formulations for drying bacterial cell extracts
AssigneeSutro Biopharma, Inc.
InventorsMartin Chandler, Kirk Hayenga, Henry Heinsohn
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Abstract
This invention provides for an improved method of stabilizing freeze dried bacterial extracts with a carbohydrate lyoprotectant such that the extracts can be for use in cell free protein synthesis. Also provided herein are formulations for stable, freeze dried bacterial extracts that when stored at room temperature retain at least about 70% protein synthesis activity compared to undried frozen bacterial extracts.
Claims19
A freeze dried bacterial extract for cell free protein synthesis comprising:
- i. dried, lysed bacterial components, wherein the extract is able to synthesize upon rehydration ATP and a target protein from a template nucleic acid encoding the target protein;
- and
- ii. a carbohydrate composition in about 0.6-2.0:1.0 (w/w) ratio of carbohydrate to dried bacterial components,
- wherein the carbohydrate composition constitutes about 22% (% of total carbohydrate composition) sucrose, 22% sorbitol, 46% mannitol, and 10% dextran, or wherein the carbohydrate composition constitutes about 100% (% of total carbohydrate composition) sucrose.
A freeze dried bacterial extract for cell free protein synthesis comprising:
- i. dried, lysed bacterial components from an Escherichia species, wherein the bacterial extract has an active oxidative phosphorylation system and, where upon rehydration the extract is able to synthesize ATP and a target protein from a template nucleic acid encoding the target protein;
- and
- ii. a carbohydrate composition in about 0.8-1.5:1.0 (w/w) ratio of carbohydrate to dried bacterial components, wherein the carbohydrate composition constitutes about 20-100% sucrose and a non-sucrose portion comprising mannitol, sorbitol, dextran or mixtures thereof.
A method for stabilizing a freeze dried bacterial extract for cell free protein synthesis, the method comprises:
- i. combining a bacterial extract comprising lysed bacterial components from an Escherichia species, wherein the bacterial extract has an active oxidative phosphorylation system and wherein the extract is able to synthesize a target protein from a template nucleic acid encoding the target protein in cell free protein synthesis, with a carbohydrate composition, wherein the carbohydrate composition constitutes about 20-100% (% of the total carbohydrate composition) sucrose and about 0-80% (% of the total carbohydrate composition) non-sucrose carbohydrate comprising mannitol, sorbitol, dextran, or mixtures thereof to yield a mixture;
- and
- ii. freeze-drying the mixture to produce the stable, freeze dried bacterial extract having an about 0.6-2:1 (w/w) ratio of carbohydrate to dried bacterial components.
Dependencies are inferred from the claim text.