Monoclonal Antibody Generation through Single B Cell Cloning
Isolation of monoclonal antibodies is an important technique for understanding the specificities and characteristics of antibodies that underlie the humoral immune response to a given antigen. Direct cloning of Ig-encoding genes from isolated B cells and their expression as monoclonal antibodies have bypassed many of these limitations and enabled the isolation of antibodies that are particularly rare or difficult to clone, such as the classic hybrodoma utilizing the Kohler and Milstein’s technique.
Oak BioSciences proudly offers high affinity monoclonal antibody developing services through our single B cell antibody technology. It offers several advantages over traditional mAb technologies, including shorter timelines, higher throughput, and compatibility with artificial intelligence (AI) powered affinity maturation. This technology is based on the analysis of the immunoglobulin gene repertoire and reactivity at the single-cell level by the application of reverse transcription-polymerase chain reaction (RT-PCR) and our expression vector cloning system. The technique involves the isolation and retrieval of antibody-coding sequences of antigen-specific B cells by leveraging miniaturization of reaction volumes. Alternatively, mAbs can be generated independently of antigen-specific B cells, comprising display technologies and, more recently, artificial intelligence-driven algorithms.
Phase 1: Single B Cell isolationthrough fluorescence-activated Cell Sorting (FACS) or other method
Phase 2: Sequencing variable region of heavy chain and light chain (VH and VL) of functional single B cell isoalted from above
Phase 3: Cloning VH and VK to Oak Biosciences vector expression cloning system
Phase 4: Monoclonal antibody expression, screening and characterization
Screening and Characterization
Assays available for antibody characterization to facilitate development:
- Western Blotting
- Endotoxin Analysis
- SEC Analysis
- IEF Gel Analysis
- Biacore Analysis
- ADCC Assays
- CDC Assays
- Epitope Mapping
1) Candidate monoclonal antibody sequences (3-10 clones)
2) Candidate monoclonal antibody protein small scale production (>100 µg)
3) Project report of candidate monoclonal antibody screening and characterization
Monoclonal Antibody Developing through Single B Cell Technologies
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