Functionalized polypeptides
View Patent ↗The invention provides functionalized polypeptides, especially therapeutic polypeptides (e.g., scFv), comprising a linker sequence that can be rapidly and specifically functionalized by the addition of one or functional moieties (e.g., PEG) or binding specificities (e.g., an amino acid sequence with a particular binding specificity). Such functionalized polypeptides are advantageous in that they have improved pharmacokinetic properties (e.g., improved in vivo half-life, tissue penetration and tissue residency time) over non-functionalized polypeptides. Methods for the rapid and reproducible generation of functionalized polypeptides are also provided.
1. A single chain antibody (scFv)comprising an antibody VH domain and an antibody VL domain that are connected by an amino acid linker; wherein the linker connecting the VH domain to the VL domain comprises two cysteines capable of forming an intrachain disulphide bond, and wherein the amino acids between the cysteines in the linker sequence form a loop when the two cysteines are disulphide bonded to one another.
2. The scFv of claim 1 , wherein the linker comprises the amino acid sequence set forth in SEQ ID No. 1.
3. The scFv of claim 1 , wherein the linker comprises the amino acid sequence set forth in SEQ ID No. 2.
4. The scFv of claim 1 , wherein the loop binds to a target molecule.
5. The scFv of claim 4 , wherein the target molecule is a PK modifier.
6. The scFv of claim 5 , wherein the PK modifier is serum albumin.
7. The scFv of claim 5 , wherein the PK modifier is a hyaluronic acid.
8. The scFv of claim 1 , wherein the linker comprises the amino acid sequence set forth in SEQ ID NO: 3.
9. The scFv of claim 1 , wherein the linker comprises the amino acid sequence set forth in SEQ ID NO: 4.
10. The scFv of claim 1 , comprising the amino acid sequence set forth in SEQ ID NO: 6 or 8.
11. A scFv of claim 1 , wherein
(a) one cysteine residue in the linker is covalently linked to a functional moiety; or
(b) both of the cysteine residues are covalently linked to a functional moiety.
12. The scFv of claim 11 , wherein the functional moiety is PEG.
13. A composition comprising the scFv of claim 1 and a pharmaceutically acceptable carrier.