Monoclonal Antibody Development

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Monoclonal Antibody Development

We combine state-of-the-art chemistry, molecular, and cell biology methods with the use of our patented DIANA technology to revolutionize monoclonal antibody development. This allows us to produce rabbit monoclonal antibodies (mAb) with unprecedented affinity and selectivity against even the most challenging targets.

Adopting our DIANA technology allows us to rapidly screen of thousands of single-antibody-containing plasma cells to identify the ones with the highest affinity and minimal cross-reactivity even before the plasma cell proceeds to sequencing and cloning. This accelerates the discovery phase but also enhances the precision of monoclonal antibody discovery.

From hapten synthesis to recombinant monoclonal antibody expression and purification

DIANA Rabbit monoclonal antibody developmentDIANA Rabbit monoclonal antibody development

Through this novel approach, we stand at the forefront of biotechnological innovation, providing clients with a new generation of antibody development services that are both robust and tailored to the advancing landscape of scientific research. The DIANA-based monoclonal antibody development is a cutting-edge technique designed to streamline the generation of high-quality, specific monoclonal antibodies. This method integrates sophisticated immunization strategies with advanced cell screening and bioengineering processes. Take a deeper view of the whole process:

  1. Immunization
    Initially, rabbits are immunized with a target antigen—this could be protein, peptide, or a small molecule. The antigen is carefully designed and prepared in-house by our experts in protein or organic chemistry departments. In the case of small molecules (or peptides), haptens are further conjugated to a carrier protein by our proprietary conjugation chemistry to boost the immune response against difficult targets. In the course of immunization, the serum is then regularly tested to ensure an optimal level of immunization, producing high titer and diverse array of selective antibodies.

  2. Cell Sorting
    Once the animal has developed desired immune response, single plasma cells, which generally carry the best antibody clones, are isolated using fluorescence-activated cell sorting (FACS). This increases the chance of finding the best performing clones from the rabbit.

  3. High-throughput screening (HTS)
    Our single cell characterization overcomes the labor and time-consuming cultivation and cloning bottle necks of the most other competing technologies. Employing DIANA’s patented high-throughput screening technology, approx. 10,000 individual plasma cells are assessed for their antibody binding properties. Key factors such as antibody affinity and cross-reactivity are measured to identify the most promising candidates for therapeutic or diagnostic applications and only the best clones proceed to sequencing and cloning. Since the amount of IgG antibodies present in a single plasma cell is sufficient for all characterization experiments in this step, no cell cultivation is needed. This is only possible due to the ridiculously high sensitivity of DIANA technology. Since plasma cells are very sensitive cells that often do not survive FACS sorting, skipping the cultivation step brings an additional advantage in the pipeline efficiency.

  4. Sequencing & Cloning
    Selected clones undergo reverse transcription-polymerase chain reaction (RT-PCR) to sequence the light and heavy chains of IgG antibody. This enables the cloning of specific antibody genes into expression vectors, ensuring that each resulting monoclonal antibody is a uniform, high-fidelity replica of the original, high-performing antibody.

  5. Expression
    Finally, these vectors are transfected into HEK293T cells, a robust expression system, where they are translated into actual antibodies. The expressed antibodies are then purified, validated, and characterized to confirm their performance.

All steps of our mAb pipeline are done in-house. Therefore, we have all steps under our control.

With our technology, we are ready to identify rare clones against challenging targets, and to measure their affinities and cross-reactivities in the early stages of our mAb pipeline.

We guarantee the highest quality.

Rabbit monoclonal antibodies offer several advantages in research and diagnostics:

Higher Affinity: Rabbit monoclonal antibodies exhibit a higher affinity for their target antigens compared to mouse mAb. Rabbit mAb can have equilibrium dissociation constant (Kd) as low as 10-13 (~ 0.1 pM). This increased binding strength also enhances their sensitivity and specificity.

Antibody Diversity: The rabbit immune system recognizes a much broader diversity of antigens compared to the mice. This diversity allows the development of antibodies that fit a wider range of epitopes, making them valuable for detecting epitopes of human origin or small molecules that are known to be non-immunogenic in mice.

Effective in Mouse Models: Rabbit monoclonal antibodies are highly desired for research on mouse models, commonly used in neuroscience, cancer, and cardiovascular studies.

Please contact us:

Vojtěch Vyklický

Head of Antibodies 

Vojtech Vyklicky Antibodies Development