Method of Supplementing Culture Media to Prevent Undesirable Amino Acid Substitutions
AssigneeBiogen Idec MA Inc.
InventorsAnurag Khetan, Yao-ming Huang, Jana Dolnikova, Nels Pederson, Helena Yusuf-Makagiansar, Paul Chen, Thomas Ryll
Life of this patent
Estimating the term…
Abstract
The present invention relates to methods of reducing substitution of amino acids during the production of polypeptides of interest in mammalian cells. By varying the concentration of amino acids in the culture medium, heterogeneity of the polypeptide's amino acid sequence is decreased, leading to higher yields of functional protein.
Claims34
A method for reducing substitution of a first amino acid by a second amino acid during translation of a polypeptide of interest in an eukaryotic cell, comprising:
- (a) culturing the cell in growth media that is supplemented with the first amino acid, or a metabolic precursor thereof, in an amount sufficient to reduce amino acid substitution;
- and/or
- (b) culturing the cell in growth media in which the amount of the second amino acid is reduced.
(canceled)
-7. (canceled)
(canceled)
-12. (canceled)
-19. (canceled)
-22. (canceled)
-26. (canceled)
-30. (canceled)
-33. (canceled)
-55. (canceled)
An antibody or fragment thereof binding to a TWEAK molecule, comprising at least 1, 2, 3, 4, 5 or 6 CDR of SEQ ID NOs 1 and 3, wherein at least one CDR comprises a serine substitution of an asparagine.
(canceled)
-65. (canceled)
A composition comprising an antibody or fragment thereof binding to a Tweak molecule, wherein less than about 10%, 5%, 3%, 1%, 0.5%, 0.3%, 0.1% of the antibodies or fragments thereof comprise at least one asparagine that is replaced by a serine.
-69. (canceled)
A method of producing an amino acid substituted library of a polypeptide of interest comprising:
- (a) analyzing the primary sequence of the polypeptide of interest to identify the amino acid to be substituted;
- (b) culturing an eukaryotic cell expressing the polypeptide of interest in media containing a decreased amount of the amino acid to be substituted;
- and
- (c) isolating the amino acid substituted polypeptides.
-84. (canceled)
Dependencies are inferred from the claim text.