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US 12,612,440 B2Granted
Modified bovine G-CSF polypeptides and their uses
AssigneesElanco US Inc., Ambrx, Inc.
InventorsAnna-Maria A. Hays Putnam, Nick Knudsen, Thea Norman, Alan Koder, Vadim Kraynov, Lillian Skidmore, Peter C. Canning
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Abstract
Modified bovine G-CSF polypeptides and uses thereof are provided.
Claims10
An isolated nucleic acid comprising at least 95% sequence identity to SEQ ID NO:
- 3, wherein the isolated nucleic acid encodes a polypeptide of SEQ ID NO:
- 1, except wherein nucleotides 397-399 of SEQ ID NO:
- 3 are substituted for an amber codon, that, when expressed with a translation system comprising an orthogonal tRNA and an orthogonal aminoacyl tRNA synthetase pair selectively incorporates a non-naturally encoded amino acid at position 133 of SEQ ID NO:
- 1.
1 more element
A method of making a bG-CSF polypeptide comprising para-acetylphenylalanine, the method comprising:
- a) culturing cells comprising a polynucleotide sequence comprising at least 95% sequence identity to
- (i) SEQ ID NO:
- 3 except at nucleotides 397-399, wherein nucleotides 397-399 of SEQ ID NO:
- 3 are substituted for an amber codon, or
- (ii) SEQ ID NO:
- 4 except at nucleotides 400-402, wherein nucleotides 400-402 of SEQ ID NO:
- 4 are substituted for an amber codon;
- in the presence of
- (i) para-acetylphenylalanine and,
- (ii) an orthogonal tRNA and aminoacyl tRNA synthetase pair capable of efficient incorporation of para-acetylphenylalanine;
- to produce a modified bG-CSF polypeptide having a para-acetylphenylalanine incorporated;
- and
- b) purifying the bG-CSF polypeptide.
10 more elements
An isolated nucleic acid comprising at least 95% sequence identity to SEQ ID NO:
- 4, wherein the isolated nucleic acid encodes a polypeptide of SEQ ID NO:
- 2, except wherein nucleotides 400-402 are substituted for an amber codon, that when expressed with a translation system comprising an orthogonal tRNA and orthogonal aminoacyl tRNA synthetase pair selectively incorporates a non-naturally encoded amino acid at position 134 of SEQ ID NO:
- 2.
Dependencies are inferred from the claim text.