CONJUGATES OF SURROGATE LIGHT CHAIN CONSTRUCTS
The present invention concerns methods for identifying, producing, and engineering binding molecule conjugates, and to the conjugates produced. In particular, the invention concerns Surrobody conjugates composed of an antibody heavy chain variable domain and a surrogate light chain, wherein the surrogate light chain and/or the antibody heavy chain variable domain is conjugated to a therapeutic or diagnostic agent.
1 . A surrogate light chain (SLC) polypeptide comprising:
a VpreB sequence and
a λ5 sequence;
wherein the VpreB sequence comprises the amino acid sequence of SEQ ID NO: 54, optionally with a cysteine residue substituted at one or more residues selected from: T16 and T21, and
wherein the λ5 sequence comprises the amino acid sequence of SEQ ID NO: 55, optionally with a cysteine residue substituted at one or more residues selected from: V60, K74, S78, V79, S85, A91, V110, V123, Q131, N133, V166, and V170.
2 . The polypeptide of claim 1 , wherein the VpreB sequence comprises at least one cysteine substitution at residues T16 or T21 or the λ5 sequence comprises at least one cysteine substitution at residues V60, K74, S78, V79, S85, A91, V110, V123, Q131, N133, V166, or V170.
3 . The polypeptide of claim 2 , wherein the VpreB sequence comprises at least one cysteine substitution at residues T16 or T21.
4 . The polypeptide of claim 2 , wherein the λ5 sequence comprises at least one cysteine substitution at residues V60, K74, S78, V79, S85, A91, V110, V123, Q131, N133, V166, or V170.
5 . A surrogate light chain (SLC) polypeptide comprising:
an amino acid sequence of SEQ ID NO: 56,
optionally with a cysteine residue substituted at one or more residues selected from: T16, T21, V107, K121, S125, V126, S132, A138, V157, V170, Q178, N180, V213, and V217.
6 . The polypeptide of claim 5 , wherein the SLC polypeptide sequence comprises at least one cysteine substitution at residues T16, T21, V107, K121, S125, V126, S132, A138, V157, V170, Q178, N180, V213, and V217.
7 . The polypeptide of claim 6 , wherein the VpreB sequence comprises at least one cysteine substitution at residues T16 or T21.
8 . The polypeptide of claim 6 , wherein the λ5 sequence comprises at least one cysteine substitution at residues V107, K121, S125, V126, S132, A138, V157, V170, Q178, N180, V213, and V217.