Polyclonal Antibody Sequencing for the Next Generation Antibody Discovery

Can you give us a brief overview of why you think it’s beneficial to generate and de novo sequence polyclonal antibodies from a single animal?
From a production point of view, it is important to have polyclonal antibodies (pAbs) that are reproducible from batch-to-batch. The standard strategy consists of mixing several pAbs isolated from different animals (2-20 animals). The reason for this is they are able to generate an averaged product in large quantities. By doing this, the pAb is less sensitive to variability.

However, we are proposing a different approach from a sequencing point of view. The previously described pAb mixture is difficult to sequence as the level of complexity is significantly increased by pooling the different animals. Instead, it would be worthwhile to assess each individual animal and select the best producer based on the unique desired quality of the generated pAbs (i.e., immunoprecipitation, Western blot, ELISA, etc.). The benefit of this approach is that you can identify the most suitable pAbs from a given producer, decode it and then make several oligo forms using recombinant protein expression. This will thereby reduce batch variability and also allow working with a product that’s better than average.



Polyclonal Antibody Sequencing for the Next Generation Antibody Discovery

What are the main challenges that arise when using polyclonal antibodies as a research tool?
Polyclonal antibodies (pAbs) are usually secreted by different B-cell clones, whereas monoclonal antibodies (mAbs) are produced by identical B-cells. What this means is that the heterogeneous nature of pAbs allows them to bind multiple epitopes of an antigen. Additionally, our native immune system generates complex pAbs in response to antigens and infection, this response is one that many scientists try to replicate in the lab. The way this response is replicated is through injecting animals with a specific antigen that will stimulate the immune response. The generated pAbs are then purified from the serum and are often pooled in order to reduce batch variability.

One of the biggest challenges of polyclonal-based development is the concept of the reproducibility crisis, which stems from batch-to-batch variability that can occur during production. The problem arises when different host animals are injected with the same antigen but then produce pAbs that have different specificities and affinities. This means that even if one was to use a new batch of antibodies, they cannot reproduce the exact same experimental results. Even more importantly, the same animal sometimes may have different immune responses to the same antigen and also exhibit time-dependent variation known as the affinity maturation process.

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