Engineered immunoglobulin heavy chain-light chain pairs and uses thereof

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.