Production and use of multimeric hemoglobins
AssigneeBaxter Biotech Technology Sarl
InventorsDavid C. Anderson, Antony J. Mathews, Gary L. Stetler
Life of this patent
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Abstract
DNA molecules which encode pseudodimeric globin-like polypeptides with an asymmetric cysteine mutation suitable for crosslinking two tetramers, or which encode pseudooligomeric globin-like polypeptides comprising four or more globin-like domains, are useful in the preparation of multimeric hemoglobin-like proteins.
Claims7
A non-naturally occurring pseudotetrameric hemoglobin-like protein comprising a pseudodimeric polypeptide having two substantially homologous globin-like domains, one of which is mutated to provide an asymmetric crosslinkable cysteine residue, wherein the corresponding residue in the other globin-like domain of said pseudodimeric polypeptide is an amino acid other than cysteine, and wherein said pseudotetramer further comprises at least one other globin-like domain-bearing polypeptide, and wherein the asymmetric crosslinkable cysteine residue is a mutation in the alpha globin-like domain corresponding to a human alpha globin position selected from the group consisting of lys7, thr8, lys11, ala12, ,gly15, lys16, gly18, ala19, his20, glu23, and thr24.
A non-naturally occurring pseudotetrameric hemoglobin-like protein comprising a pseudodimeric polypeptide having two substantially homologous globin-like domains, one of which is mutated to provide an asymmetric crosslinkable cysteine residue, wherein the corresponding residue in the other globin-like domain of said pseudodimeric polypeptide is an amino acid other than cysteine, and wherein said pseudotetramer further comprises at least one other globin-like domain-bearing polypeptide, and wherein the asymmetric crosslinkable cystine residue is a mutation in the beta globin-like domain corresponding to a human beta globin position selected from the group consisting of his2, leu3, thr4, glu6, ser9, thr12, ala13, gly16, lys17, val18, asn19, val20, asp21, and glu22.
A recombinant mutant hemoglobin-like protein comprising a mutation in a beta-globin like domain at amino acid position 93, wherein the cysteine at position 93 is replaced with an amino acid chosen from the group consisting of alanine and threonine.
Dependencies are inferred from the claim text.