Engineered immunoglobulin heavy chain-light chain pairs and uses thereof
Provided are 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.
1. A set of polynucleotides encoding an antibody construct comprising a first heterodimer and a second heterodimer,
the 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 the 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, and the set of polynucleotides comprises:
a) a first polynucleotide encoding H1 comprising amino acid substitutions 146G and 179K, a second polynucleotide encoding L1 comprising amino acid substitutions 124E, 160E and 180E, a third polynucleotide encoding H2 comprising amino acid substitutions 143E and 145T, and a fourth polynucleotide encoding L2 comprising amino acid substitutions 160K and 178R, or conservative substitutions thereof;
b) a first polynucleotide encoding H1 comprising amino acid substitutions 143K and 146G, a second polynucleotide encoding L1 comprising amino acid substitutions 124E and 133D, a third polynucleotide encoding H2 comprising amino acid substitutions 143E and 145T, and a fourth polynucleotide encoding L2 comprising amino acid substitution 124R, or conservative substitutions thereof;
c) a first polynucleotide encoding H1 comprising amino acid substitutions 146G and 179R, a second polynucleotide encoding L1 comprising amino acid substitutions 124E, 160E and 178D, a third polynucleotide encoding H2 comprising amino acid substitutions 145T, 179D, and 188L, and a fourth polynucleotide encoding L2 comprising amino acid substitutions 160K and 178R, or conservative substitutions thereof;
d) a first polynucleotide encoding H1 comprising amino acid substitutions 143A, 146G and 179R, a second polynucleotide encoding L1 comprising amino acid substitutions 124E, 133W, 160E and 180E, a third polynucleotide encoding H2 comprising amino acid substitutions 145T, 179D, and 188F, and a fourth polynucleotide encoding L2 comprising amino acid substitutions 133A, 160K and 178R, or conservative substitutions thereof; or
e) a first polynucleotide encoding H1 comprising amino acid substitutions 146G and 186R, a second polynucleotide encoding L1 comprising amino acid substitutions 124E, 160E and 178D, a third polynucleotide encoding H2 comprising amino acid substitutions 145E, 146G, 179D, and 188L, and a fourth polynucleotide encoding L2 comprising amino acid substitutions 124R, 160K and 178R, or conservative substitutions thereof.
2. The set of polynucleotides according to claim 1 , wherein L1 and/or L2 are kappa light chains.
3. The set of polynucleotides according to claim 1 , wherein the first heterodimer and the second heterodimer each comprise a full-length IgG heavy chain having an Fc domain.
4. The set of polynucleotides according to claim 3 , wherein the Fc domain of H1 interacts preferentially with the Fc domain of H2 as compared to forming a homodimer.
5. The set of polynucleotides according to claim 1 , wherein H1 and H2 are from an IgG1, IgG2, IgG3, or IgG4 antibody.
6. The set of polynucleotides according to 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 set of polynucleotides according to claim 1 , wherein the thermal stability as measured by the melting temperature (Tm) of at least one of the first and second heterodimers is within about 10° C. of the Tm of the corresponding heterodimer without the amino acid substitutions.
8. The set of polynucleotides according to 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 set of polynucleotides according to claim 1 , wherein the antibody construct is bispecific.
10. One or more vectors comprising the set of polynucleotides according to claim 1 .
11. A prokaryotic or eukaryotic host cell comprising the one or more vectors according to claim 10 .
12. A method for preparing the antibody construct according to claim 1 comprising the steps of:
transforming a host cell with one or more vectors comprising any one of (a) to (e);
culturing the host cell under conditions that allow expression of the antibody construct; and
recovering the antibody construct from the culture.