AMINO ACID SUBSTITUTED MOLECULES
First Claim
1. A method for producing a modified target polypeptide, comprising(a) providing a host cell, the host cell comprising a vector having a polynucleotide encoding the target polypeptide,(b) site-specifically incorporating one or more non-natural amino acid codons into the polynucleotide, wherein at least one non-natural amino acid codon corresponds to the first position of the amino terminus of the target polypeptide,(c) growing the host cell under conditions such that the host cell expresses the target polypeptide, wherein the target molecule retains the non-natural amino acid residue at the first position of the amino terminus, and wherein the non-natural amino acid residue at the first position of the amino terminus contains an azide, alkyne, vinyl, or aryl halide group, thereby producing a modified target polypeptide.
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Accused Products
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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Citations
90 Claims
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1. A method for producing a modified target polypeptide, comprising
(a) providing a host cell, the host cell comprising a vector having a polynucleotide encoding the target polypeptide, (b) site-specifically incorporating one or more non-natural amino acid codons into the polynucleotide, wherein at least one non-natural amino acid codon corresponds to the first position of the amino terminus of the target polypeptide, (c) growing the host cell under conditions such that the host cell expresses the target polypeptide, wherein the target molecule retains the non-natural amino acid residue at the first position of the amino terminus, and wherein the non-natural amino acid residue at the first position of the amino terminus contains an azide, alkyne, vinyl, or aryl halide group, thereby producing a modified target polypeptide.
- 14. A composition comprising a modified target polynucleotide encoding a target polypeptide, the target polynucleotide comprising one or more non-natural amino acid codons wherein at least one non-natural amino acid codon contains an azide, alkyne, vinyl, or aryl halide group and corresponds to the first position of the amino terminus of the target polypeptide.
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26. A pharmaceutical composition comprising a modified target polypeptide comprising a target polypeptide having one or more non-natural amino acids residues incorporated, wherein at least one of the non-natural amino acid residues corresponds to the first position of the amino terminus of the target polypeptide.
- 27. A method for modifying a target molecule comprising altering one or more wild type amino acid residues to another naturally occurring amino acid residue, and altering one or more wild type amino acid residues to a non-natural amino acid residue.
- 36. A modified target molecule comprising one or more wild type amino acid residues altered to another naturally occurring amino acid residue, one or more wild type amino acid residues altered to a non-natural amino acid residue, and a chemical moiety attached to the non-natural amino acid.
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45. A kit comprising at least one reagent for modifying a target molecule, and instructions for modifying the molecule according to a method comprising altering one or more wild type amino acid residue to another naturally occurring amino acid residue, and altering one or more wild type amino acid residue to a non-natural amino acid residue.
- 46. A method for modifying a molecule comprising substituting one or more amino acid residues with a non-natural amino acid residue and adding a chemical moiety to said non-natural amino acid residue by a cross-coupling technique, wherein said molecule retains a native function.
- 69. A modified target molecule comprising one or more amino acid residues substituted with a non-natural amino acid residue, wherein a chemical moiety is coupled to said non-natural amino acid residue, and wherein the modified target molecule retains a native function.
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90. A kit comprising at least one reagent for modifying a target molecule, and instruction for modifying said molecule according to a method comprising substituting one or more amino acid residues with a non-natural amino acid residue and adding one or more chemical moieties to said molecule.
Specification