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US 12,624,377 B2Granted
Systems and method for the production of Griffithsin and related proteins
AssigneeDuke University
InventorsMichael Lynch, John Decker
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Abstract
Methods and kits are provided for producing Griffithsin. The methods include providing a genetically modified microorganism comprising a gene encoding Griffithsin protein operably linked to an inducible promotor and growing the genetically modified microorganism under conditions that induce the promotor and cause expression of griffithisin. The Griffithsin is purified by releasing Griffithsin from the microorganism by cellular disruption, performing a precipitation step to remove contaminating protein and nucleic acids, and performing an anion exchange chromatography step.
Claims10
A method of producing Griffithsin comprising:
- i) providing a genetically modified E. coli microorganism comprising a gene encoding a Griffithsin protein operably linked to an inducible promotor;
- ii) growing the genetically modified microorganism under conditions that induce the promotor and cause expression of Griffithsin, the inducible promotor expresses the Griffithsin protein when phosphate is depleted from media in which the genetically modified microorganism is growing;
- iii) releasing Griffithsin from the microorganism by cellular disruption;
- iv) performing a precipitation step to remove contaminating protein and nucleic acids;
- v) performing an anion exchange chromatography step,
- wherein the precipitation step and anion exchange chromatograph step produce purified Griffithsin, and
- wherein the Griffithsin is a protein of the red algae Griffithsia.
4 more elements
Dependencies are inferred from the claim text.