Publications
bioRxivApr 2024 DOI:
10.1101/2024.04.22.589762

De novodesign of high-affinity single-domain antibodies

van der Kant, Rob; Zhang, Zhongyao; Marković, Iva; Vizarraga, David; Garcia, Teresa; Maragkou, Katerina; Blanco, Javier Delgado; Cianferoni, Damiano; Orlando, Gabriele; Carolis, Carlo; Volkov, Alexander N.; Savvides, Savvas N.; Dewilde, Maarten; Schymkowitz, Joost; Pubul, Luis Serrano; Rousseau, Frederic
Product Used
Oligo Pools
Abstract
Developing antibodies is complex and resource-intensive, and methods for designing antibodies targeting specific epitopes are lacking. We introduce ade novoantibody design approach leveraging the empirical force field FoldX to design complementarity determining regions (CDRs). Starting from a scaffold VHH, we tackled three challenges of increasing difficulty: 1) design the CDRs to optimize VHH stability and affinity for its original target; 2) design the CDRs for high affinity to the human ortholog; 3) design the CDRs for low nanomolar affinity for a pre-defined epitope on the unrelated human Interleukin-9 receptor alpha, for which no antibodies were previously developed. For each challenge we reached single digit nanomolar affinity in a single design cycle. Our approach allowsde novodesign of high-affinity VHHs while ensuring specificity and stability.
Product Used
Oligo Pools

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