Production of Human Antibody Fragments Binding to Melittin and Phospholipase A2 in Africanised Bee Venom: Minimising Venom Toxicity

Abstract

The hybrid created from the crossbreeding of European and African bees, known as the Africanised bee, has provided numerous advantages for current beekeeping. However, this new species exhibits undesirable behaviours, such as colony defence instinct and a propensity to attack en masse, which can result in serious accidents. To date, there is no effective treatment for cases of Africanised bee envenomation. One promising technique for developing an efficient antivenom is the use of phage display technology, which enables the production of human antibodies, thus avoiding the complications of serum therapy, such as anaphylaxis and serum sickness. The aim of this study was to produce human monoclonal single-chain Fv (scFv) antibody fragments capable of inhibiting the toxic effects of Africanised bee venom. We conducted four rounds of selection of antibodies against the venom and three rounds of selection of antibodies against purified melittin. Three clones were selected and tested by enzyme-linked immunosorbent assay to verify their specificity for melittin and phospholipase A2. Two clones (C5 and C12) were specific for melittin, and one (A7) was specific for phospholipase A2. In a kinetic haemolytic assay, these clones were evaluated individually and in pairs. The A7-C12 combination had the best synergistic effect and was chosen to be used in the assays of myotoxicity inhibition and lethality. The A7-C12 combination inhibited the in vivo myotoxic effect of the venom and increased the survival of treated animals.

Engineering Venom’s Toxin-Neutralizing Antibody Fragments and Its Therapeutic Potential

“…Here, we focus on the most recent advances in the design of recombinant antibodies against snake and scorpion toxins. We also know that occasional studies have been undertaken for developing antidotes against other venoms [54]. …”

Massive attack of honeybee on macaws ( Ara ararauna and Ara chloropterus ) in Brazil – A case report

“…Brazil may be considered the cradle of the hybrid Africanized honeybee since African bees (Apis mellifera scutellata) imported in the 1950s were cross-bred with European species (Apis mellifera and Apis mellifera ligustica). The crossbreeding resulted in a bee that produces more honey than the African bee but displays more aggressive behavior than the European species, and this has led to various types of accidents with severe and even fatal results for both animals and humans (Almeida et al, 2011;Clarke et al, 2002;Ferreira et al, 2012;Funayama et al, 2012). This hybrid spread quickly throughout South America, mainly due to the high reproductive rate and migratory behavior of the species, which has increased the number of accidents involving bees (Ferreira et al, 2012).…”

Scorpion Venoms

“…Advancements in antivenom optimization have been reached not only in scorpion issues but also against other medically important poisonous animals such as snakes (Chavanayarn et al 2012), spiders (Bugli et al 2008), and bees (Funayama et al 2012).…”