Mutants of human interleukin-3
AssigneeDsm N.V.
InventorsLambertus Christiaan Johannes Dorssers, Robert Willem van Leen
Life of this patent
Estimating the term…
Abstract
Biologically active deletion and substitution mutants of hIL-3 are provided. Preferred mutants are those having one or more deletions at the N-terminus (amino acids 1-14) and/or the C-terminus (amino acids 116-133, 120-130 and/or 130-133). Preferred substitution mutants include Cys16 →Ala16 and/or Cys84→Ala84, Glu50→Lys50 and Lys79 →Glu79. These mutants can be used to formulate pharmaceutical compositions. Also disclosed are antibodies directed against specific epitopes localized between amino acids 29 and 54.
Claims18
A human interleukin-3 antagonist having an amino acid sequence which differs from the sequence of human interleukin-3 by the replacement of one or both of Cys16 or Cys84 by another amino acid so as to disturb the disulfide bridge between Cys16 or Cys84.
A human interleukin-3 antagonist having an amino acid sequence which differs from the sequence of human interleukin-3 by a charge reversal substitution at one or more of positions 36, 50, and 79.
A human interleukin-3 antagonist having an amino acid sequence which differs from the sequence of human interleukin-3 by the substitutions Arg63Ala64→Pro63Gly64.
A method to produce a biologically active analog of human interleukin-3 wherein said analog differs from the amino acid sequence of human interleukin-3 by having deleted therefrom 4 to 18 amino acids from the C terminus of human interleukin-3, which method comprises
- culturing cells transformed with a vector, comprising a DNA sequence encoding a polypeptide analog of human interleukin-3 having the amino acid sequence of human interleukin-3 having deleted therefrom 4 to 18 amino acids from the C terminus of the native human interleukin-3 sequence, in combination with transcription and translation regulation sequences that enable the expression of said polypeptide analog,
- under conditions suitable for producing said analog;
- and
- recovering the analog from the culture.
Dependencies are inferred from the claim text.