Rabbit B-Cell Cloning Robust Methodology for Antibody Discovery

The Fusion Antibodies Process

• Isolation of B-cells – Peripheral blood, bone marrow, or lymphoid tissue samples are collected, and antigen-specific B-cells are identified using flow cytometry or single-cell sorting.
• Amplification of antibody genes – The immunoglobulin heavy and light chain genes from individual B-cells are cloned using molecular biology sequencing techniques.
• Reconstruction and Ab-ility™ Screening – The full IgG sequences are screened using our Ab-ility™ in-silico developability tool to determine best candidates for expression.
• Recombinant Expression – The selected sequences are expressed in recombinant systems (e.g. mammalian cells) to produce full monoclonal antibodies for further testing.
• Screening and characterization – The resulting antibodies are evaluated for binding affinity, specificity, stability and any other client specific functional properties.

Fig 1. Fusion Antibodies B-Cell Cloning Process

Benefits as a Discovery Technology

• Greater Epitope Diversity – Rabbits produce antibodies that recognise epitopes often missed by mice, including small molecules, post-translational modifications, and highly conserved or hidden epitopes.
• Higher Affinity Antibodies – Rabbit immune repertoires undergo more extensive somatic hypermutation, often yielding antibodies with low- and even sub-nanomolar affinities that surpass those typically generated by murine hybridomas.
• Direct Access to Natural VH/VL Pairing – B-cell cloning preserves the native heavy- and light-chain pairing from single rabbit B-cells, ensuring antibodies maintain their natural functional properties. Murine hybridomas can sometimes mis-pair during fusion or lose rare clones.

B-cell cloning leverages the natural immune system’s ability to generate highly effective antibodies, making it one of the most powerful and clinically relevant discovery technologies for modern antibody therapeutics.

All antibodies developed using this technology can be humanized using our CDRx® Platform and we have success rate of 100% in various species.

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