Engineered immunoglobulin heavy chain-light chain pairs and uses thereof

A computer-readable storage medium storing

• a dataset comprising data representing complementary mutations in a first heterodimer comprising a first immunoglobulin heavy chain polypeptide sequence (H1) and a first immunoglobulin light chain polypeptide sequence (L1);
• and a second heterodimer comprising a second immunoglobulin heavy chain polypeptide sequence (H2) and a second immunoglobulin light chain polypeptide sequence (L2), wherein H1 and H2 each comprise at least a heavy chain variable domain (VH domain) and a heavy chain constant domain (CH1 domain);
• wherein L1 and L2 each comprise at least a light chain variable domain (VL domain) and a light chain constant domain (CL domain), and wherein the dataset of complementary mutations comprises data representing those mutations listed in Tables 29, 30 or 31 or a subset of those mutations;
• and.
• computer executable code for determining the likelihood that H1 will pair preferentially with L1 as compared to L2 and/or H2 will pair preferentially with L2 as compared to L1.

A computer implemented method for determining preferential pairing, comprising:

• obtaining a dataset comprising data representing complementary mutations in a first heterodimer comprising a first immunoglobulin heavy chain polypeptide sequence (H1) and a first immunoglobulin light chain polypeptide sequence (L1);
• and a second heterodimer comprising a second immunoglobulin heavy chain polypeptide sequence (H2) and a second immunoglobulin light chain polypeptide sequence (L2), wherein H1 and H2 each comprise at least a heavy chain variable domain (VH domain) and a heavy chain constant domain (CH1 domain);
• wherein L1 and L2 each comprise at least a light chain variable domain (VL domain) and a light chain constant domain (CL domain), and wherein the dataset of complementary mutations comprises data representing those mutations listed in Tables 29, 30 or 31 or a subset of those mutations;
• and
• determining, by a computer processor, the likelihood that H1 will pair preferentially with L1 as compared to L2 and/or H2 will pair preferentially with L2 as compared to L1.

A method of producing a bi-specific antigen binding polypeptide construct, said bi-specific construct comprising a first heterodimer comprising a first immunoglobulin heavy chain polypeptide sequence (H1), and a first immunoglobulin light chain polypeptide sequence (L1) from a first mono-specific antigen binding polypeptide;

• and a second heterodimer comprising a second immunoglobulin heavy chain polypeptide sequence (H2), and a second immunoglobulin light chain polypeptide sequence (L2) from a second mono-specific antigen binding polypeptide, wherein H1 and H2 each comprise at least a heavy chain variable domain (VH domain) and a heavy chain constant domain (CH1 domain);
• wherein L1 and L2 each comprise at least a light chain variable domain (VL domain) and a light chain constant domain (CL domain), the method comprising:
• a. obtaining a dataset comprising data representing a set of amino acid modifications within H1, H2, L1 and L2 such that upon introduction of a subset of the modifications into H1, H2, L1 and/or L2, H1 pairs preferentially with L1 as compared to L2 and H2 pairs preferentially with L2 as compared to L1 in a test system;
• b. introducing a subset of one or more modifications from the dataset into the first heterodimer and/or the second heterodimer;
• and
• c. co-expressing the first heterodimer and the second heterodimer in at least one host cell to produce an expression product comprising the bi-specific construct.