US 8,541,203 B2 · Activatable binding polypeptides and methods of identification and use thereof

Abstract

Activatable binding polypeptides (ABPs), which contain a target binding moiety (TBM), a masking moiety (MM), and a cleavable moiety (CM) are provided. Activatable antibody compositions, which contain a TBM containing an antigen binding domain (ABD), a MM and a CM are provided. Furthermore, ABPs which contain a first TBM, a second TBM and a CM are provided. The ABPs exhibit an “activatable” conformation such that at least one of the TBMs is less accessible to target when uncleaved than after cleavage of the CM in the presence of a cleaving agent capable of cleaving the CM. Further provided are libraries of candidate ABPs, methods of screening to identify such ABPs, and methods of use. Further provided are ABPs having TBMs that bind VEGF, CTLA-4, or VCAM, ABPs having a first TBM that binds VEGF and a second TBM that binds FGF, as well as compositions and methods of use.

Claims41

Dependent claims

A method of manufacturing an activatable binding polypeptide (ABP), the method comprising:

(a) culturing a cell comprising a nucleic acid construct that encodes the ABP under conditions that lead to expression of the ABP, wherein the ABP comprises a masking moiety (MM), a cleavable moiety (CM), and an antigen binding domain (ABD) that specifically binds vascular endothelial growth factor (VEGF),
(i) wherein the ABP in an uncleaved state comprises a structural arrangement from N-terminus to C-terminus as follows:
MM-CM-ABD or ABD-CM-MM;
(ii) wherein the MM is a peptide that inhibits binding of the ABD to the target, and wherein the MM comprises an amino acid sequence selected from the group consisting of SEQ ID NOs:
31, 32, 33, 34, 35, 36, 37 and 38;
and
(iii) wherein, the CM is positioned in the ABP such that, in an uncleaved state, the MM interferes with specific binding of the ABD to VEGF, and in a cleaved state the MM does not interfere or compete with specific binding of the ABD to VEGF;
(b) recovering the ABP;
and
(c) testing the ABP for the ability to maintain an activatable phenotype while in soluble form.

7 more elements

Dependencies are inferred from the claim text.

Activatable binding polypeptides and methods of identification and use thereof

Life of this patent

Activatable binding polypeptides and methods of identification and use thereof

Life of this patent

Abstract

Claims41