TY - JOUR PY - 2011// TI - Production of Human Antibody Fragments Binding to Melittin and Phospholipase A2 in Africanised Bee Venom: Minimising Venom Toxicity JO - Basic and Clinical Pharmacology and Toxicology A1 - Funayama, Jaqueline C. A1 - Pucca, Manuela B. A1 - Roncolato, Eduardo Crosara A1 - Bertolini, Thaís Barboza A1 - Campos, Lucas B. A1 - Barbosa, José E. SP - 290 EP - 297 VL - 110 IS - 3 N2 - 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 ELISA 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 use 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.

Language: en

LA - en SN - 1742-7835 UR - http://dx.doi.org/10.1111/j.1742-7843.2011.00821.x ID - ref1 ER -