Engineered immunoglobulin heavy chain-light chain pairs and uses thereof

Patent metadata

Field	Value
Publication number	US-9914785-B2
Application number	US-201314092804-A
Country	US
Kind code	B2
Filing date	Nov 27, 2013
Priority date	Nov 28, 2012
Publication date	Mar 13, 2018
Grant date	Mar 13, 2018

Abstract

The present invention provides heterodimer pairs comprising a first heterodimer and a second heterodimer wherein each heterodimer comprises an immunoglobulin heavy chain or fragment thereof and an immunoglobulin light chain. At least one of the heterodimers comprises amino acid modifications in the C H1 and/or C L domains, amino acid modifications in the V H and/or V L domains or a combination thereof. The modified amino acid residues are part of the interface between the light chain and heavy chain and are modified in order to create preferential pairing between each heavy chain and a desired light chain such that when the two heavy chains and two light chains of the heterodimer pair are co-expressed in a mammalian cell, the heavy chain of the first heterodimer preferentially pairs with one of the light chains rather than the other. Likewise, the heavy chain of the second heterodimer preferentially pairs with the second light chain rather than first.

First claim

We claim:

1. An antibody construct comprising at least a first heterodimer and a second heterodimer,
said first heterodimer comprising a first human or humanized immunoglobulin G (IgG) heavy chain polypeptide (H1) and a first human or humanized immunoglobulin light chain polypeptide (L1), and said second heterodimer comprising a second human or humanized IgG heavy chain polypeptide (H2) and a second human or humanized immunoglobulin light chain polypeptide (L2),
H1 and H2 each comprising a heavy chain variable domain (VH domain) and a heavy chain constant domain 1 (CH1 domain), and L1 and L2 each comprising a light chain variable domain (VL domain) and a light chain constant domain (CL domain);
wherein H1, H2, L1 and L2 comprise the following amino acid substitutions at positions identified according to the Kabat numbering system:
a) H1 comprises amino acid substitutions 146G and 179K, L1 comprises amino acid substitutions 124E, 160E and 180E, H2 comprises amino acid substitutions 143E and 145T, and L2 comprises amino acid substitutions 160K and 178R, or conservative substitutions thereof;
b) H1 comprises amino acid substitutions 143K and 146G, L1 comprises amino acid substitutions 124E and 133D, H2 comprises amino acid substitutions 143E and 145T, and L2 comprises amino acid substitution 124R, or conservative substitutions thereof;
c) H1 comprises amino acid substitutions 146G and 179R, L1 comprises amino acid substitutions 124E, 160E and 178D, H2 comprises amino acid substitutions 145T, 179D, and 188L, and L2 comprises amino acid substitutions 160K and 178R, or conservative substitutions thereof;
d) H1 comprises amino acid substitutions 143A, 146G and 179R, L1 comprises amino acid substitutions 124E, 133W, 160E and 180E, H2 comprises amino acid substitutions 145T, 179D, and 188F, and L2 comprises amino acid substitutions 133A, 160K and 178R, or conservative substitutions thereof; or
e) H1 comprises amino acid substitutions 146G and 186R, L1 comprises amino acid substitutions 124E, 160E and 178D, H2 comprises amino acid substitutions 145E, 146G, 179D, and 188L, and L2 comprises amino acid substitutions 124R, 160K and 178R, or conservative substitutions thereof;
and wherein the amino acid substitutions promote preferential pairing of H1 with L1 as compared to L2, and/or of H2 with L2 as compared to L1, when H1, H2, L1 and L2 are co-expressed in a cell.

2. The antibody construct of claim 1 , wherein L1 and/or L2 are kappa light chains.

3. The antibody construct of claim 1 , wherein said first heterodimer and said second heterodimer each comprise a full-length IgG heavy chain having an Fc domain.

4. The antibody construct of claim 3 , wherein the Fc domain of H1 interacts preferentially with the Fc domain of H2 as compared to forming a homodimer.

5. The antibody construct of claim 1 , wherein H1 and H2 are from an IgG1, IgG2, IgG3, or IgG4 antibody.

6. The antibody construct of claim 1 , wherein when both L1 and L2 are co-expressed with at least one of H1 and H2, the relative yield of the at least one of H1-L1 and H2-L2 heterodimer pair to that of the corresponding mispaired H1-L2 or H2-L1 heterodimer pair is greater than 50%.

7. The antibody construct of claim 1 , wherein the thermal stability as measured by the melting temperature (Tm) of at least one of said first and second heterodimers is within about 10° C. of the Tm of the corresponding heterodimer without the amino acid substitutions.

8. The antibody construct of claim 1 , wherein the affinity of each heterodimer for its antigen is within about 50-fold of the affinity of the wild type heterodimer for the antigen.

9. The antibody construct of claim 1 , wherein the antibody construct is bispecific.

10. An antibody construct comprising at least a first heterodimer and a second heterodimer,
said first heterodimer comprising a first human or humanized immunoglobulin G (IgG) heavy chain polypeptide (H1) and a first human or humanized immunoglobulin light chain polypeptide (L1), and said second heterodimer comprising a second human or humanized IgG heavy chain polypeptide (H2) and a second human or humanized immunoglobulin light chain polypeptide (L2),
H1 and H2 each comprising a heavy chain variable domain (VH domain) and a heavy chain constant domain 1 (CH1 domain), and L1 and L2 each comprising a light chain variable domain (VL domain); and a light chain constant domain (CL domain);
wherein H1, H2, L1 and L2 comprise the following amino acid substitutions at positions identified according to the Kabat numbering system:
a) H1 comprises amino acid substitutions 146G and 179K, L1 comprises amino acid substitutions 124E, 160E and 180E, H2 comprises amino acid substitutions 143E and 145T, and L2 comprises amino acid substitutions 160K and 178R, or conservative substitutions thereof;
b) H1 comprises amino acid substitutions 143K and 146G, L1 comprises amino acid substitutions 124E and 133D, H2 comprises amino acid substitutions 143E and 145T, and L2 comprises amino acid substitution 124R, or conservative substitutions thereof;
c) H1 comprises amino acid substitutions 146G and 179R, L1 comprises amino acid substitutions 124E, 160E and 178D, H2 comprises amino acid substitutions 145T, 179D, and 188L, and L2 comprises amino acid substitutions 160K and 178R, or conservative substitutions thereof;
d) H1 comprises amino acid substitutions 143A, 146G and 179R, L1 comprises amino acid substitutions 124E, 133W, 160E and 180E, H2 comprises amino acid substitutions 145T, 179D, and 188F, and L2 comprises amino acid substitutions 133A, 160K and 178R, or conservative substitutions thereof; or
e) H1 comprises amino acid substitutions 146G and 186R, L1 comprises amino acid substitutions 124E, 160E and 178D, H2 comprises amino acid substitutions 145E, 146G, 179D, and 188L, and L2 comprises amino acid substitutions 124R, 160K and 178R, or conservative substitutions thereof.

11. A composition comprising a set of polypeptides comprising a first human or humanized immunoglobulin light chain polypeptide (L1), a second human or humanized immunoglobulin light chain polypeptide (L2), L1 and L2 each comprising a light chain constant domain (CL domain) and a light chain variable domain (VL domain), and a human or humanized immunoglobulin G (IgG) heavy chain polypeptide (H1) comprising a heavy chain variable domain (VH domain) and a heavy chain constant domain 1 (CH1 domain), wherein H1, L1 and L2 comprise the following amino acid substitutions at positions identified according to the Kabat numbering system:
a) H1 comprises amino acid substitutions 143A, 146G, and 179R, L1 comprises amino acid substitutions 124E, 133W, 160E, and 180E, and L2 comprises amino acid substitutions 133A, 160K, and 178R, or conservative substitutions thereof;
b) H1 comprises amino acid substitutions 143E and 145T, L1 comprises amino acid substitutions 124R, and L2 comprises amino acid substitutions 124E and 133D, or conservative substitutions thereof;
c) H1 comprises amino acid substitutions 143E and 145T, L1 comprises amino acid substitutions 160K and 178R, and L2 comprises amino acid substitutions 124E, 160E, and 180E, or conservative substitutions thereof;
d) H1 comprises amino acid substitutions 143K and 146G, L1 comprises amino acid substitutions 124E and 133D, and L2 comprises amino acid substitutions 124R, or conservative substitutions thereof;
e) H1 comprises amino acid substitutions 145E, 146G, 179D, and 188L, L1 comprises amino acid substitutions 124R, 160K, and 178R, and L2 comprises amino acid substitutions 124E, 160E, and 178D, or conservative substitutions thereof;
f) H1 comprises amino acid substitutions 145T, 179D, and 188F, L1 comprises amino acid substitutions 133A, 160K,

Assignees

• Zymeworks Inc

Inventors

• Corper Adam Louis
• Urosev Dunja
• Tom-Yew Stacey A L
• Bleile Dustin Weyland Blue
• Von Kreudenstein Thomas Spreter
• Dixit Surjit
• Lario Paula Irene
• Sanches Mario

Classifications

• C07K2317/55

  Fab or Fab' · CPC title

• C07K2317/94

  Stability, e.g. half-life, pH, temperature or enzyme-resistance · CPC title

• C07K16/36 Primary

  against blood coagulation factors · CPC title