Antigenic GM-CSF peptides and antibodies to GM-CSF

Inventors

Sass, Philip M. • Nicolaides, Nicholas C. • Grasso, Luigi • Ebel, Wolfgang • Li, Jian • Chao, Qimin • Routhier, Eric

Assignees

Eisai Inc

Abstract

Hybridoma lines that secrete human monoclonal antibodies with high binding specificity and biological activity, particularly neutralizing activity against granulocyte-macrophage colony stimulating factor, and methods of generating the hybridoma lines are provided. Target antigens and epitopes are also provided. The antibodies may be used in therapeutic methods, for example in the treatment of cancer, infectious disease, or autoimmune disease.

Core Innovation

The patent describes human anti-GM-CSF immunotherapeutics based on hybridoma-derived monoclonal antibodies that specifically bind to granulocyte-macrophage colony stimulating factor (GM-CSF). It states that high-affinity, human, biologically active neutralizing GM-CSF antibodies are desired. The disclosure associates the antibodies with specific GM-CSF antigenic peptide epitopes and provides epitope mapping of GM-CSF regions using overlapping GM-CSF peptides.

The patent describes an approach for generating hybridomas and screening to obtain human monoclonal antibodies with GM-CSF specificity and neutralization. It further discusses optimizing hybridoma diversity and stability using mismatch repair (MMR) inhibition, including morphocenes/anthracene MMR inhibitors, followed by clone selection. Candidate antibodies are identified (E5/E10/G9 and G7), including performance characterization such as neutralization in TF-1 GM-CSF-dependent bioassays and binding affinity measurements (e.g., BIACORE KD values), together with stability/production metrics.

The disclosure provides sequence identification and definitions for the GM-CSF antigenic peptides (SEQ ID NOs:1-5, including mature GM-CSF regions) and for epitope peptide regions (SEQ ID NOs:35-39). It also specifies antibody sequence information, including antibody CDRs (heavy/light CDR1-3 and optional framework regions) and associated nucleic acid and accession information for deposited antibodies and/or cells. The patent supplies the sequence framework underlying antibodies defined by heavy-chain and light-chain CDR sequences that correspond to the cited SEQ ID NOs and to deposited cells (e.g., ATCC Accession Nos. PTA-8173 and PTA-8193).

Claims Coverage

The independent claims cover recombinant polynucleotides, isolated cells, and isolated hybridoma cells that recombinantly express or encode GM-CSF-binding antibodies, with the antibodies defined either by explicit heavy/light CDR sequence identities (six CDRs using SEQ ID NOs) or by the six complementarity determining regions of an antibody produced by a deposited cell (ATCC Accession No. PTA-8173). Across the independent claims, the inventive concepts focus on binding specificity to GM-CSF combined with defined CDR composition, applied to polynucleotides and multiple cellular formats.

Overall, the claim set provides coverage for GM-CSF-binding antibodies in which the antibody identity is anchored by either explicit heavy and light chain CDR sequences (SEQ ID NOs:40-45) or by the six complementarity determining regions of an antibody produced by a specific deposited cell (ATCC Accession No. PTA-8173). This CDR-defined antibody identity is then implemented across recombinant polynucleotides, isolated cells that recombinantly express the antibody, and isolated hybridoma cells that express the monoclonal antibody.

Stated Advantages

Documented Applications