Ge Healthcare Bioprocess R&d Ab

Ge Healthcare Bio-sciences Ab

Cytiva Bioprocess R&d Ab

Goran Bauren

Gustav Jose Rodrigo

Tomas Bjorkman

Mats Ander

Per-Mikael Aberg

The invention discloses a polypeptide with improved alkaline stability, which polypeptide comprises a mutant of a B or C domain ofProtein A (SpA), as specified by SEQ ID NO 1 or SEQ ID NO 2, or of Protein Z, as specified by SEQ ID NO 3, wherein at least the glutamine residue at position 9 has been mutated to an amino acid other than asparagine. The invention also discloses multimers of said polypeptide, as well as separation matrices comprising the multimers or polypeptides.

Mutated immunoglobulin-binding polypeptides

The invention discloses an immunoglobulin-binding protein comprising one or more mutated immunoglobulin-binding domains (monomers) of staphylococcal Protein A (E, D, A, B, C) or protein Z or a functional variant thereof, wherein in at least one of the one or more mutated monomers, the asparagine or histidine at the position corresponding to H18 of the B domain of Protein A or of Protein Z has been deleted or substituted with a first amino acid residue which is not proline or asparagine and wherein, if the amino acid residue at position 57 is proline and the amino acid residue at position 28 is asparagine, then the amino acid residue at the position corresponding to H18 of the B domain of protein A or of protein Z is not serine, threonine or lysine.

Affinity chromatography matrix

The invention discloses a polypeptide with improved alkaline stability, which polypeptide comprises a mutant of a B or C domain of Staphylococcus Protein A (SpA), as specified by SEQ ID NO 1 or SEQ ID NO 2, or of Protein Z, as specified by SEQ ID NO 3, wherein at least the glutamic acid residue at position 15 has been mutated to an amino acid other than asparagine. The invention also discloses multimers of said polypeptide, as well as separation matrices comprising the multimers or polypeptides.

A polypeptide with improved alkaline stability, which polypeptide comprises a mutant of a B or C domain ofProtein A, as specified by SEQ ID NO 1 or SEQ ID NO 2, or of Protein Z, as specified by SEQ ID NO 3, wherein at least the glutamine residue at position 9 has been mutated to an amino acid other than asparagine. The invention also discloses multimers of said polypeptide, as well as separation matrices comprising the multimers or polypeptides.

A polypeptide with improved alkaline stability, which polypeptide comprises a mutant of a B or C domain ofProtein A, as specified by SEQ ID NO 1 or SEQ ID NO 2, or of Protein Z, as specified by SEQ ID NO 3, wherein at least the glutamine residue at position 15 has been mutated to an amino acid other than asparagine. The invention also discloses multimers of the polypeptide, as well as separation matrices comprising the multimers or polypeptides.

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Uppsala, Sweden

Goran Bauren