Cross-reactivity and inhibition myotoxic effects induced by Bothrops snake venoms using specific polyclonal anti-BnSP7 antibodies

• Published in: Biologicals 2017-11, Vol.50, p.109-116
• Main Authors: Melo, Lamartine L., Mendes, Mirian M., Alves, Lívia M., Isabel, Thais F., Vieira, Sâmela A.P.B., Gimenes, Sarah N.C., Soares, Andreimar M., Rodrigues, Veridiana M., Izidoro, Luiz F.M.
• Format: Article
• Language: English
• Subjects: BnSP-7-PLA2; Bothrops pauloensis; Cross-reactivity; Inhibition; Phospholipase A2; Polyclonal antibodies
• ISSN: 1045-1056; 1095-8320
• DOI: 10.1016/j.biologicals.2017.08.002

Polyclonal antibodies raised in Balb-c mice against BnSP-7, a Lys-49 phospholipase A2, were used to measure cross reactivity against other snake venoms. Using ELISA, these antibodies were able to recognize PLA2s isoforms present in venoms of botropic snakes at 1:6400, 1:3200 and 1:100 ratios (w/w). These antibodies highly recognized proteins of low molecular weight (∼14,000) from crude snake venom Bp and Bm by Western Blotting. PLA2 these venoms, by alignment of primary structures demonstrated high identity with BnSP-7 PLA2, especially in the C-terminal region. However, the crude snake venom Bd and Bj, showed low recognition. The PLA2 activity of Bothrops pauloensis, Bothrops moojeni venoms or BpPLA2-TXI was inhibited significantly when anti-BnSP-7 antibodies were incubated at 1:10 and 1:20 ratios (venoms or toxin:anti-BnSP-7, w/w), respectively. The myotoxic effect induced by the same venoms was also reduced significantly at 1:1, 1:10 and 1:20 ratios, by decreased creatine kinase levels. The anti-PLA2 polyclonal antibodies effectively recognized PLA2s from Bothrops pauloensis and Bothrops moojeni venoms, and neutralized specific catalytic and myotoxic activity.