Enhancing the Bothropic Antivenom through a Reverse Antivenomics Approach

Antivenoms are the only effective treatment for snakebite envenomation and have saved countless lives over more than a century. Despite their value, antivenoms present risks of adverse reactions. Current formulations contain a fraction of nonspecific antibodies and serum proteins. While new promising candidates emerge as the next generation of antivenoms, it remains clear that animal-derived antivenoms will still play a critical role for years to come. In this study, we improved the bothropic antivenom (BAv), by capturing toxin-specific antibodies through affinity chromatography using immobilized <i>Bothrops jararaca</i> venom toxins. This process produced an improved antivenom (iBAv) enriched in neutralizing antibodies and depleted of serum proteins. Proteomic analysis showed that iBAv was 87% depleted in albumin and 37-83% lower in other serum proteins compared to BAv. Functional evaluation demonstrated that iBAv had a 2.9-fold higher affinity for venom toxins by surface plasmon resonance and a 2.8-fold lower ED50 <i>in vivo</i>, indicating enhanced potency. Our findings indicate that enriching specific antibodies while depleting serum proteins reduces the total protein dose required and increases the potency of antivenom. Although technical and economic considerations remain for large-scale implementation, this affinity-enriched antivenom represents a significant advancement in improving antivenom efficacy against <i>B. jararaca</i> envenomations.