A single-chain variable fragment selected against a conformational epitope of a recombinantly produced snake toxin using phage display

• Published in: New biotechnology 2023-09, Vol.76, p.23-32
• Main Authors: Rimbault, Charlotte, Knudsen, Pelle D., Damsbo, Anna, Boddum, Kim, Ali, Hanif, Hackney, Celeste M., Ellgaard, Lars, Bohn, Markus-Frederik, Laustsen, Andreas H.
• Format: Article
• Language: English
• Subjects: Animals; antibodies; Antibodies, Monoclonal; Antibody selection; bacteriophages; Bacteriophages - metabolism; biotechnology; cholinergic receptors; Epitopes; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Monoclonal antibodies; peptide libraries; Peptide Library; Phage display; Recombinant toxins; Single-Chain Antibodies - genetics; Snake toxins; Snake Venoms - metabolism; Snakebite envenoming; snakes; toxins; venoms
• ISSN: 1871-6784; 1876-4347
• DOI: 10.1016/j.nbt.2023.04.002

Phage display technology is a powerful tool for selecting monoclonal antibodies against a diverse set of antigens. Within toxinology, however, it remains challenging to generate monoclonal antibodies against many animal toxins, as they are difficult to obtain from venom. Recombinant toxins have been proposed as a solution to overcome this challenge, but so far, few have been used as antigens to generate neutralizing antibodies. Here, we describe the recombinant expression of α-cobratoxin in E. coli and its successful application as an antigen in a phage display selection campaign. From this campaign, an scFv (single-chain variable fragment) was isolated with similar binding affinity to a control scFv generated against the native toxin. The selected scFv recognizes a structural epitope, enabling it to inhibit the interaction between the acetylcholine receptor and the native toxin in vitro. This approach represents the first entirely in vitro antibody selection strategy for generating neutralizing monoclonal antibodies against a snake toxin. •Functional alpha-cobratoxin can be recombinantly expressed in E. coli.•Recombinant α-cobratoxin can be used as antigen for phage display selection.•Selected scFv has a binding affinity on a par with previously discovered scFvs.•The scFv binds to a structural epitope of the α-cobratoxin.