ALBUMIN VARIANTS BINDING TO FCRN
The invention relates to methods of identifying albumin variants having improved pharmacokinetics, albumin variants having improved pharmacokinetics, and therapeutic uses of albumin variants having improved pharmacokinetics.
1 . A method of identifying a human serum albumin (HSA) variant, the method comprising
a. providing a mutated HSA; and
b. determining whether the mutated HSA has at least one mutation in domain III that decreases fatty acid binding compared to fatty acid binding by a wild type HSA, wherein a mutated HSA that decreases fatty acid binding compared to a wild type HSA is an HSA variant.
2 . The method of claim 1 , further comprising
c. determining the binding affinity of the mutated HSA for FcRn,
wherein a mutated HSA that can bind to FcRn with the same or increased affinity compared to binding of a wild type HSA to FcRn is an HSA variant.
3 . The method of claim 1 , further comprising
d. determining the PK of the mutated HSA compared to the PK of a wild type HSA,
wherein a mutated HSA that has increased PK compared to a wild type HSA is an HSA variant.
4 . A human serum albumin (HSA) variant comprising at least one mutation in domain III that decreases fatty acid binding to the HSA variant compared to fatty acid binding by a wild type HSA.
5 . The HSA variant of claim 4 , wherein the HSA variant can bind to FcRn.
6 . The HSA variant of claim 4 , wherein the HSA variant has an increased PK compared to a wild type HSA.
7 . The HSA variant of claim 4 , wherein the mutation
a. alters one or more residues in domain III of a wild type HSA that can bind to a carboxyl; or
b. alters one or more residues in domain III that are lining residues.
8 . The HSA variant of claim 7 , wherein the HSA variant is mutated at one or more residues selected from the group consisting of R410, Y411, S489, Y401, and K525.
9 . The HSA variant of claim 8 , wherein the residue is mutated to a non-polar amino acid or a negatively charged amino acid.
10 . The HSA variant of claim 9 , wherein the mutated residue is an alanine or glutamic acid.
11 . The HSA variant of claim 7 , wherein the mutated residue is a lining residue selected from the group consisting of Y411, V415, V418, T422, L423, V426, L430, L453, L457, L460, V473, R485, F488, L491, F502, F507, F509, K525, A528, L529, L532, V547, M548, F551, L575, V576, S579, and L583.
12 . The HSA variant of claim 11 , wherein the mutated residue is selected from the group consisting of Y411, V415, V418, L423, V426, L430, L453, L457, L460, V473, P485, F488, L491, F502, F507, F509, A528, L529, L532, V547, M548, F551, L575, V576, and L583.
13 . The HSA variant of claim 12 , wherein the mutated residue is mutated to a serine.
14 . An HSA variant covalently linked to a therapeutic agent.
15 . A method of identifying a scaffold molecule, the method comprising
a. providing a candidate molecule; and
b. determining whether the candidate molecule can bind to an HSA and can inhibit fatty acid binding to the HSA,
wherein, a candidate molecule that can bind to an HSA and can inhibit fatty acid binding is a scaffold molecule.
16 . The method of claim 15 , wherein the scaffold molecule can bind to one or more of residues R410, Y411, S489, Y401, or K525 of a wild type HSA.
17 . A scaffold molecule identified by the method of claim 15 .
18 . The scaffold molecule identified by the method of claim 17 further comprising a therapeutic molecule, thereby forming a heterogeneous scaffold molecule, wherein the PK of the heterogeneous scaffold molecule is increased compared to the PK of the therapeutic molecule.
19 . A method of increasing the serum half-life of a molecule, the method comprising covalently linking the molecule to the HSA variant of claim 4 .
20 . The method of claim 19 , wherein the molecule is a protein or polypeptide.
21 . A molecule comprising the HSA variant of claim 4 and a heterologous molecule.
22 . The molecule of claim 21 , wherein the heterologous molecule is a protein or polypeptide.