Monovalent interleukin 12 (il-12) heterodimeric fc proteins

A method for producing a monovalent IL-12 heterodimeric Fc protein, comprising:

• (a) providing a first nucleic acid molecule encoding a first polypeptide comprising an IL-12p35 subunit fused to the N-terminus of a first immunoglobulin fragment crystallizable (Fc) region and a second nucleic acid molecule encoding a second polypeptide comprising an IL-12p40 subunit fused to the N-terminus of a second Fc region, wherein the first Fc region and the second Fc region each comprise one or more amino acid substitutions that promote heterodimerization, and wherein the IL-12p35 subunit of the monovalent IL-12 heterodimer Fc protein comprises one or more amino acid substitutions that reduce the potency of the monovalent IL-12 heterodimeric Fc protein compared to a monovalent IL-12 heterodimeric Fc protein comprising wild-type IL-12p35 and IL-12p40 subunits;
• (b) transfecting a host cell with the first nucleic acid molecule and the second nucleic acid molecule to produce a transfected host cell;
• (c) cultivating the host cell in a culture medium under conditions for the production of the first polypeptide and the second polypeptide, which are excreted into the culture medium and associated into a monovalent IL-12 heterodimeric Fc protein comprising a first polypeptide and a second polypeptide;
• and
• (d) recovering the monovalent IL-12 heterodimeric Fc proteins from the culture medium.

A polypeptide comprising the amino acid sequence set forth in SEQ ID NO:

• 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 13, 14 15, 16 17, 18, 19, 20, 21, 22, 23, 24 25 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, or 66;
• or the amino acid sequence set forth in SEQ ID NO:
• 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, or 132;
• or the amino acid sequence set forth in SEQ ID NO:
• 133, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, or 198;
• or the amino acid set forth in SEQ ID NO:
• 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 278, 279, 280, 281, 282, or 283;
• or the amino acid set forth in SEQ ID NO:
• 284, 285, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, or 303;
• or the amino acid set forth in SEQ ID NO:
• 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, or 326;
• or the amino acid set forth in SEQ ID NO:
• 327, 328, 329, 330, 331, 332, 333, 334, 335,336, 337, 338, 339, 340, 341, 342, 343, 344, 345, or 346;
• or the amino acid set forth in SEQ ID NO:
• 347, 348, 349, 350, 351, 352, 353, 354, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, or 369;
• or the amino acid set forth in SEQ ID NO:
• 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, or 389.

A nucleic acid molecule comprising the nucleotide sequence set forth in SEQ ID NO:

• 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 249, or 250;
• or the nucleotide sequence set forth in SEQ ID NO:
• 234, 235, 236, 237, 240, 241, 242, 243, 244, 245, 246, 247, or 248;
• or the nucleotide sequence set forth in SEQ ID NO:
• 251, 252, 253, 254, 255, 256, 257, 258, 259, or 260.