Immunoglobulin binding polypeptides
AssigneeNavigo Proteins GmbH
InventorsHanna Bobolowski, Erik Fiedler, Ulrich Haupts
Life of this patent
Estimating the term…
Abstract
The present invention relates to novel engineered artificial immunoglobulin (Ig) binding polypeptides comprising three domains and two linkers. The invention further relates to affinity matrices comprising these artificial Ig binding molecules of the invention. The novel Ig binding proteins are particularly useful for the affinity purification of proteins requiring elution at a higher pH (in particular higher than pH 4.2). The invention also relates to a use of the novel Ig binding proteins or affinity matrices for affinity purification of immunoglobulins and to methods of affinity purification using the novel Ig binding proteins of the invention.
Claims14
An immunoglobulin (Ig)-binding protein comprising the following general structure from N- to C-terminus:
- domain 1-linker 1-domain 2-linker 2-domain 3, wherein:
- (i) domain 1 comprises an amino acid sequence, which is at least 70% identical to the amino acid sequence of SEQ ID NO:
- 1 (SQSANLLAEAKKLNDAQ) or to SEQ ID NO:
- 4 (SVSLEILAEAKKLNDAQ);
- (ii) linker 1 has at least 3 and no more than 5 amino acid residues, and further wherein linker 1 comprises the amino acid sequence:
- (a) X1LT or X1IT, wherein X1 is selected from the group consisting of glycine (G), arginine (R), asparagine (N), proline (P), alanine (A), aspartic acid (D), glutamic acid (E), and threonine (T);
- or
- (b) GGIX2 (SEQ ID NO:
- 19) or GGLX2 (SEQ ID NO:
- 20), wherein X2 is selected from the group consisting of threonine (T) and serine(S);
- or
- (c) GGX3LT (SEQ ID NO:
- 83), wherein X3 is selected from the group consisting threonine (T) and serine(S);
- or
- (d) X4GLT (SEQ ID NO:
- 84), wherein X4 is selected from the group consisting of arginine (R) and glycine (G) and asparagine (N);
- or
- (e) GX5LT (SEQ ID NO:
- 85), wherein X5 is selected from the group consisting of arginine (R) and glycine (G) and asparagine (N);
- or
- (f) an amino acid sequence selected from the group consisting of GGITT (SEQ ID NO:
- 106), NGLT (SEQ ID NO:
- 107), RGLT (SEQ ID NO:
- 108), GRLT (SEQ ID NO:
- 109), and PTL;
- (iii) domain 2 comprises an amino acid sequence, which is at least 70% identical to the amino acid sequence of SEQ ID NO:
- 8 (EEQRNGFIQSLKDD) or to SEQ ID NO:
- 9 (EEQRNAFIQSLRDD);
- (iv) linker 2 has at least 6 and no more than 8 amino acid residues;
- and
- (v) domain 3 comprises an amino acid sequence, which is at least 70% identical to the amino acid sequence of SEQ ID NO:
- 13 (NKEQQNAFYEILHL) or to SEQ ID NO:
- 15 (DKEQQAAFYEILHL).
Dependencies are inferred from the claim text.