A new immunochemistry process that transform a non-immunogenic crotamine-like antigen from rattlesnake (Crotalus durissus cumanensis) venom, in immunogenic to produce anti-crotamine-like antibodies

Abstract

The making of antibodies in animals can be demanding due to that several antigens, mostly of low molecular masses, provoke imperceptible immune response or are even totally nonimmunogenic. The transformation of non-immunogenic molecules into effective antigens represent an important immunological tasks. The crotamine from the rattlesnake Crotalus durissus cumanensis snake venom was purified by a Mono S HR 10/10 chromatography column and used to immunise C57/B mice, after to be polymerised with glutaraldehyde. The murine polyclonal antibodies directed against native crotamine-like (NCL) treated with glutaraldehyde and their product crotamine-like polymer (CLP) were generated by immunisation injecting CLP via lymph node cells. These antibodies were capable of detecting CLP in an enzyme-linked immunosorbent assay. The SDS-PAGE of NCL and CLP showed bands of molecular masses ~ 3 kDa and ~18 kDa, respectively. These results offer evidence that the polyclonal antibodies recognise specific putative original and post-polymerisation epitopes on the CLP molecule, which were maintained following the process of polymerisation. The results are discussed in relation to the preservation of a functional postpolymerisation epitopes on CLP.