Animal-derived antivenoms constitute the mainstay in the therapy of snakebite envenoming. The efﬁcacy of antivenoms to neutralize toxicity of medically-relevant snake venoms has to be demonstrated through meticulous preclinical testing before their introduction into the clinical setting. The gold standard in the preclinical assessment and quality control of antivenoms is the neutralization of venom-induced lethality. In addition, depending on the pathophysiological proﬁle of snake venoms, the neutralization of other toxic activities has to be evaluated, such as hemorrhagic, myotoxic, edema-forming, dermonecrotic, in vitro coagulant, and deﬁbrinogenating effects. There is a need to develop laboratory assays to evaluate neutralization of other relevant venom activities. The concept of the 3Rs (Replacement, Reduction, and Reﬁnement) in Toxinology is of utmost importance, and some advances have been performed in their implementation. A signiﬁcant leap forward in the study of the immunological reactivity of antivenoms against venoms has been the development of “antivenomics”, which brings the analytical power of mass spectrometry to the evaluation of antivenoms. International partnerships are required to assess the preclinical efﬁcacy of antivenoms against snake venoms in different regions of the world in order to have a detailed knowledge on the neutralizing proﬁle of these immunotherapeutics.