Capitalizing on the ability to alternate antigens, we carried out several cross-panning strategies to enrich for cross-reactive binders. As an in vitro antibody discovery method, phage display selection allows for a high level of control during the discovery process, such as the possibility to alter pH, alternate antigens, and reduce the antigen concentration, to mention a few 6, 11. In this study, we explored the use of antibody phage display technology to isolate cross-reactive single-chain variable fragment (scFv) antibodies. For the discovery of cross-reactive antibodies, phage display is a key technology 6, 7, which can be applied together with specific methods such as cross-panning 3, 8, 9, and/or next-generation sequencing analysis of parallel phage display pannings 10. This is especially important for diseases with endogenous targets, such as autoimmune diseases and cancer. However, cross-reactivity towards protein isoforms from different animal species is also of particular relevance for translational aspects between preclinical models and the clinical setting. These traits are especially relevant for developing therapies against indications such as infectious diseases and envenomings. Additionally, antibodies can be developed to be cross-reactive and have broad toxin-neutralizing capabilities 3, 4, 5, given a proper discovery strategy. However, when antigens share the (exact) same functions, this seems to increase the chances of selecting cross-reactive antibodies, which may possibly be due to the existence of structurally similar motifs on the antigens.Īntibodies have become a highly successful group of therapeutic molecules in recent decades due to their ability to bind antigens with high selectivity and specificity 1, 2. Further, we find that the feasibility of discovering cross-reactive antibodies using cross-panning cannot easily be predicted by analyzing the sequence, structural, or surface similarity of the antigens alone.
We showcase how cross-panning can increase the chances of discovering cross-reactive single-chain variable fragments (scFvs) from phage display campaigns. Therefore, we sought to explore how a previously reported phage display-based cross-panning strategy drives the selection of cross-reactive antibodies using seven different snake toxins belonging to three protein (sub-)families: phospholipases A 2, long-chain α-neurotoxins, and short-chain α-neurotoxins. However, the mechanisms driving antibody cross-reactivity typically remain to be elucidated. Such antibodies have been successfully selected against closely related antigens using phage display technology. Antibodies with cross-reactive binding and broad toxin-neutralizing capabilities are advantageous for treating indications such as infectious diseases and animal envenomings.
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