Amino acid substituted molecules
First Claim
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1. A method for producing a modified target polypeptide,wherein the modified target polypeptide includes at least one non-natural amino acid and retains the function of the corresponding wild-type target polypeptide, the method comprising:
- providing a host cell comprising a heterologous polynucleotide that encodes the target polypeptide, wherein the polynucleotide has been mutated such that;
a region or location where a post-translational modification is desired in the target polypeptide comprises at least one codon for a selected amino acid; and
at least one codon for the selected amino acid is substituted with a codon for a different amino acid outside the region or location where a post-translational modification is desired, wherein the modified target polypeptide retains function; and
growing the host cell in a medium containing the non-natural amino acid under conditions sufficient for the host cell to incorporate the non-natural amino acid into at least one position of the selected amino acid in the region or location where a post-translational modification is desired in the target polypeptide,thereby producing a modified target polypeptide that comprises a non-natural amino acid in the region or location where post-translational modification is desired.
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Abstract
The invention provides compositions and methods of identifying, modifying and producing modified target molecules, including therapeutic molecules by modification with non-natural amino acids. Certain aspects of the invention include methods of adding a chemical moiety to a target molecule, and the compositions resulting therefrom. Certain aspects of the invention also relate to kits for identifying, modifying and producing modified target molecules described herein.
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29 Claims
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1. A method for producing a modified target polypeptide,
wherein the modified target polypeptide includes at least one non-natural amino acid and retains the function of the corresponding wild-type target polypeptide, the method comprising: -
providing a host cell comprising a heterologous polynucleotide that encodes the target polypeptide, wherein the polynucleotide has been mutated such that; a region or location where a post-translational modification is desired in the target polypeptide comprises at least one codon for a selected amino acid; and at least one codon for the selected amino acid is substituted with a codon for a different amino acid outside the region or location where a post-translational modification is desired, wherein the modified target polypeptide retains function; and growing the host cell in a medium containing the non-natural amino acid under conditions sufficient for the host cell to incorporate the non-natural amino acid into at least one position of the selected amino acid in the region or location where a post-translational modification is desired in the target polypeptide, thereby producing a modified target polypeptide that comprises a non-natural amino acid in the region or location where post-translational modification is desired. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29)
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Specification