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AssigneeJanssen Biotech, Inc.
InventorsMark Anderson, Ricardo Attar, Michael Diem, Linus Hyun, Alastair King, Steven Jacobs, Shalom Goldberg, Donna Klein, Sheri Moores, Karyn O'Neil, Kristen Picha
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Cysteine engineered monospecific and bispecific EGFR and/or c-Met FN3 domain containing molecules comprising one or more free cysteine amino acids are prepared by mutagenizing a nucleic acid sequence of a parent molecule and replacing one or more amino acid residues by cysteine to encode the cysteine engineered FN3 domain containing monospecific or bispecific molecules; expressing the cysteine engineered FN3 domain containing molecules; and recovering the cysteine engineered FN3 domain containing molecule. Isolated cysteine engineered monospecific or bispecific FN3 domain containing molecules may be covalently attached to a detection label or a drug moiety and used therapeutically.
1) An isolated cysteine engineered fibronectin type III (FN3) domain comprising at least one cysteine substitution at a position selected from the group consisting of residues 6, 8, 10, 11, 14, 15, 16, 20, 30, 34, 38, 40, 41, 45, 47, 48, 53, 54, 59, 60, 62, 64, 70, 88, 89, 90, 91, and 93 of the FN3 domain, wherein the FN3 domain is based on SEQ ID NO:
3) An isolated cysteine engineered fibronectin type III (FN3) domain comprising the amino acid sequence of SEQ ID NO:
4) An isolated cysteine engineered fibronectin type III (FN3) domain comprising the amino acid sequence of SEQ ID NO:
7) A method of preparing a cysteine engineered FN3 domain comprising:
18) An isolated cysteine engineered fibronectin type III (FN3) domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:
19) An isolated cysteine engineered bispecific FN3 molecule comprising a first fibronectin type III (FN3) domain and a second FN3 domain, wherein the first FN3 domain comprises a cysteine substitution at a position selected from the group consisting of residues 6, 8, 10, 11, 14, 15, 16, 20, 30, 34, 38, 40, 41, 45, 47, 48, 53, 54, 59, 60, 62, 64, 70, 88, 89, 90, 91, and 93 of the first FN3 domain, specifically binds epidermal growth factor receptor (EGFR) and blocks binding of epidermal growth factor (EGF) to EGFR, and the second FN3 domain specifically binds hepatocyte growth factor receptor (c-Met), and blocks binding of hepatocyte growth factor (HGF) to c-Met.
31) A method of preparing an isolated cysteine engineered bispecific molecule comprising:
35) The method of 34, further comprising a step selected from the group consisting of:
36) An isolated cysteine engineered fibronectin type III (FN3) domain comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:
37) An isolated cysteine engineered bispecific FN3 molecule comprising a first fibronectin type III (FN3) domain and a second FN3 domain, wherein the first FN3 domain comprises a cysteine substitution at a position selected from the group consisting of residues 7, 9, 11, 12, 15, 16, 17, 21, 31, 35, 39, 41, 42, 46, 48, 49, 54, 55, 60, 61, 63, 65, 71, 89, 90, 91, 92, and 94 of the first FN3 domain, specifically binds epidermal growth factor receptor (EGFR) and blocks binding of epidermal growth factor (EGF) to EGFR, and the second FN3 domain specifically binds hepatocyte growth factor receptor (c-Met), and blocks binding of hepatocyte growth factor (HGF) to c-Met.
39) Any invention described herein.
Dependencies are inferred from the claim text.