Modified variable domain molecules and methods for producing them b
AssigneeGarvan Institute of Medical Research
InventorsDaniel Christ, Kip Dudgeon, Romain Rouet
Life of this patent
Estimating the term…
Abstract
The present disclosure provides an isolated, engineered or non-naturally occurring protein comprising an antibody light chain variable domain (VL) which may comprise at least one negatively charged amino acid positioned between residues 49 to 56 according to the numbering system of Kabat, the protein capable of binding specifically to an antigen.
Claims36
An isolated, engineered or non-naturally occurring protein comprising:
- a) an antibody light chain variable domain (VL) comprising a negatively charged amino acid at two or more positions between residues 49 and 56 according to the numbering system of Kabat, the protein capable of specifically binding to an antigen;
- or
- b) (i) an antibody light chain variable domain (VL) comprising a negatively charged amino acid at one or more positions between residues 49 and 56 according to the numbering system of Kabat;
- and
- (ii) an antibody heavy chain variable domain (VH) comprising a negatively charged amino acid at one or more positions selected from the group consisting of residues 28, 30, 31, 32, 33 and 35 according to the numbering system of Kabat, wherein the protein is capable of specifically binding to an antigen;
- or
- c) a modified antibody light chain variable domain (VL) capable of specifically binding to an antigen, wherein the VL comprises a negatively charged amino acid at one or more positions selected from the group consisting of residues 49, 51, 52, 53 and 56 according to the numbering system of Kabat, and wherein the unmodified form of the VL does not comprise the negatively charged amino acid(s);
- or
- d) a modified antibody light chain variable domain (VL) capable of specifically binding to an antigen, wherein the VL comprises a negatively charged amino acid at two or more positions between residues 49 and 56 according to the numbering system of Kabat, and wherein the unmodified form of the VL does not comprise two or more negatively charged amino acid(s) at the positions;
- or
- e) (i) a modified antibody light chain variable domain (VL) comprising a negatively charged amino acid at a position between residues 49 and 56 according to the numbering system of Kabat, wherein the unmodified form of the VL does not comprise a negatively charged amino acid at the position;
- and
- (ii) a modified antibody heavy chain variable domain (VH) comprising a negatively charged amino acid at one or more positions selected from the group consisting of residues 28, 30, 31, 32, 33 and 35 according to the numbering system of Kabat, wherein the unmodified form of the VH does not comprise a negatively charged amino acid at the position, wherein the modified protein is capable of specifically binding to an antigen.
(canceled)
11-14. (canceled)
16-17. (canceled)
19-20. (canceled)
A library including proteins comprising antibody light chain variable domains (VLs), the VLs comprising negatively charged amino acids at one or more positions between residues 49 and 56 according to the numbering system of Kabat.
A method for increasing the aggregation-resistance of a protein comprising an antibody light chain variable domain (VL), the method comprising:
- a) modifying the VL by substituting an amino acid at one or more positions selected from the group consisting of residues 49, 50, 51, 52, 53 and 56 according to the numbering system of Kabat with a negatively charged amino acid;
- or
- b) modifying the VL such that it comprises negatively charged amino acids at two or more positions between residues 49 and 56 according to the numbering system of Kabat, wherein the unmodified protein does not comprise the two or more negatively charged amino acids;
- or
- c) when the protein further comprises an antibody heavy chain variable domain (VH), modifying the protein such that it comprises:
- (i) a negatively charged amino acid at one or more positions between residues 49 and 56 of the VL according to the numbering system of Kabat;
- and
- (ii) a negatively charged amino acid at one or more positions selected from the group consisting of residues 28, 30, 31, 32, 33 and 35 of the VH according to the numbering system of Kabat, wherein the protein prior to modification does not comprise a negatively charged amino acid at the positions in the VL and the VH.
32-33. (canceled)
(canceled)
Dependencies are inferred from the claim text.