Anti-EGFR monoclonal antibody, methods of making, and uses thereof in the treatment of cancer
AssigneeShanghai Biomabs Pharmaceuticals Co., Ltd.
InventorWeizhu Qian
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Abstract
The present invention provides a method for producing an anti-EGFR monoclonal antibody and the applications thereof. The method comprises the steps of: designing and synthesizing the light chain and heavy chain according to the codons preferred by Chinese hamster, transfecting GS knockout host cells CHO-CR-GS−/−, culturing cells using serum-free technology, isolating and purifying the antibody, and obtaining the low immunogenicity CMAB009 antibody.
Claims42
A composition comprising an antibody, and a pharmaceutically acceptable carrier,
- wherein the antibody comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO:
- 2 and a heavy chain comprising an amino acid sequence set forth in SEQ ID NO:
- 4, and
- wherein the antibody has a z-average (z-avg) of 10-25 nm as determined by a dynamic light scattering (DLS) analysis,
- wherein the Fab segments of the antibody have a sialic acid NANA glycan chain structure, and
- wherein the antibody does not comprise an N-glycolylneuraminic acid (NGNA) and does not comprise a Gal-α(1,3)-Gal glycan,
- wherein the antibody binds epidermal growth factor receptor (EGFR).
A liquid pharmaceutical composition comprising water and an anti-EGFR antibody, wherein the anti-EGFR antibody comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO:
- 2 and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:
- 4,
- wherein the antibody has a z-average (z-avg) of 10-25 nm as determined by dynamic light scattering (DLS) analysis, and the anti-EGFR antibody does not comprise an N-glycolylneuraminic acid (NGNA), and does not comprise a Gal-α(1,3)-Gal glycan.
A liquid pharmaceutical composition comprising water and an anti-EGFR antibody,
- wherein the anti-EGFR antibody comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO:
- 2 and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:
- 4,
- wherein the anti-EGFR antibody is produced in a Chinese Hamster Ovary (CHO) cell,
- and wherein the composition does not comprise a polysorbate and/or a saccharobiose.
A liquid pharmaceutical composition consisting essentially of water, an anti-EGFR antibody, sodium chloride, sodium dihydrogen phosphate dihydrate, and disodium phosphate dihydrate,
- wherein the anti-EGFR antibody comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO:
- 2 and a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:
- 4, and
- wherein the anti-EGFR antibody does not comprise an N-glycolylneuraminic acid (NGNA) glycan, does not comprise a Gal-α(1,3)-Gal glycan, and/or does comprise a Gal-α(2, 3/6)-Gal glycan.
A method of producing an anti-EGFR monoclonal antibody, said method comprising:
- a) an anti-EGFR monoclonal antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO:
- 2 and a heavy chain comprising the amino acid sequence of SEQ ID NO:
- 4;
- b) constructing a recombinant plasmid using a nucleic acid encoding the light chain and the heavy chain;
- c) transfecting CHO cells with the recombinant plasmid;
- d) screening for a high-expressing clone;
- e) culturing in large scale, the high-expressing clone, wherein a cell culture temperature is 33° C.˜36° C., a pH of a cell culture growth media is 6.5˜6.9, and an osmotic pressure of the cell culture growth media is 290 mOsm/kg˜350 mOsm/kg;
- f) isolating the cultured clone of e);
- and
- g) purifying the isolated clone of f) to obtain the anti-EGFR monoclonal antibody.
A method of inhibiting progression of cancer in a human subject, said method comprising administering the composition of claim to the subject, such that progression is inhibited.
A method of treating or inhibiting progression of colorectal cancer in a subject having colorectal cancer, said method comprising administering an anti-EGFR antibody and irinotecan, such that colorectal cancer is treated, wherein the antibody comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO:
- 2, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:
- 4, and contains a Gal-α 2, 3/6)-Gal glycan.
A method of treating or inhibiting progression of colorectal cancer in a subject having colorectal cancer, said method comprising administering an anti-EGFR antibody and irinotecan, such that colorectal cancer is treated, wherein the antibody comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO:
- 2, comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO:
- 4, and does not contain either an N-glycolylneuraminic acid (NGNA) glycan or a Gal-α(1,3)-Gal glycan.
Dependencies are inferred from the claim text.