Antibodies that bind CD3 epsilon

Inventors

Sato, Aaron • Garg, Pankaj • Liu, Qiang • YUAN, Tom

Assignees

Twist Bioscience Inc • Twist Bioscience Corp

Abstract

Provided herein are methods and compositions relating to variant nucleic acid libraries encoding for antibodies including single domain antibodies. Libraries generated using methods described herein have improved characteristics including improved binding affinity. Libraries described herein include variegated libraries comprising nucleic acids each encoding for a predetermined variant of at least one predetermined reference nucleic acid sequence. Further described herein are protein libraries generated when the nucleic acid libraries are translated. Further described herein are cell libraries expressing variegated nucleic acid libraries described herein.

Core Innovation

The invention relates to an antibody or antibody fragment that binds to CD3 epsilon (CD3E). The antibody or antibody fragment comprises a variable region of a heavy chain (VH) with CDRH1, CDRH2, and CDRH3, and a variable region of a light chain (VL) with CDRL1, CDRL2, and CDRL3. The CDRH1/CDRH2/CDRH3 and CDRL1/CDRL2/CDRL3 comprise amino acid sequences corresponding to defined SEQ ID NO sets, including SEQ ID NOs 118, 121, 124, 129, 132, and 135, or 119, 122, 125, 130, 133, and 136, or 120, 123, 126, 131, 134, and 137.

The described framework includes antibody variant libraries and selection approaches for generating CD3 epsilon-binding antibodies and fragments. It specifies variegated and precise codon-variant nucleic acid libraries and selection and characterization based on binding performance using affinity thresholds such as KD <100 nM.

The disclosure also provides extensive sequence definitions and identity/homology relationships for CDRH1/CDRH2/CDRH3 and CDRL1/CDRL2/CDRL3 relative to defined SEQ ID NOs associated with CD3 epsilon. It further discusses platform aspects for highly parallel nucleic acid library synthesis and a downstream expression/assay workflow to evaluate binding performance.

Claims Coverage

Independent claim covers CD3 epsilon-binding antibodies or antibody fragments with a defined heavy-chain CDR set and a defined light-chain CDR set, where the CDR amino-acid sequences correspond to one of three SEQ ID NO sets. Dependent claims further refine the heavy-chain and/or light-chain variable regions by specific sequences or identity thresholds, and extend coverage to isolated nucleic-acid embodiments encoding the antibody.

Overall, the claims are directed to a CD3 epsilon-binding antibody or antibody fragment defined by specific CDRH1/CDRH2/CDRH3 and CDRL1/CDRL2/CDRL3 amino-acid sequences, with dependent claims narrowing to particular heavy- and light-chain variable-region sequence requirements, including identity thresholds, and extending coverage to isolated nucleic-acid embodiments encoding the antibody.

Stated Advantages

Documented Applications