artículo científico
High-density peptide microarray exploration of the antibody response in a rabbit immunized with a neurotoxic venom fraction
Fecha
2017-11Registro en:
0041-0101
10.1016/j.toxicon.2017.08.028
28867663
Autor
Engmark, Mikael Gerling
Jespersen, Martin Closter
Lomonte, Bruno
Lund, Ole
Laustsen, Andreas Hougaard
Institución
Resumen
Polyvalent snakebite antivenoms derive their therapeutic success from the ability of their antibodies to
neutralize venom toxins across multiple snake species. This ability results from a production process
involving immunization of large mammals with a broad suite of toxins present in venoms. As a result of
immunization with this wide range of toxins, many polyvalent antivenoms have a high degree of crossreactivity
to similar toxins in other snake venoms e a cross-reactivity which cannot easily be deconvoluted.
As a proof of concept, we aimed at exploring the opposite scenario by performing a highthroughput
evaluation of the extent of cross-reactivity of a polyclonal mixture of antibodies that was
raised against only a single snake venom fraction. For this purpose, a venom fraction containing short
neurotoxin 1 (SN-1; Uniprot accession number P01416, three-finger toxin (3FTx) family), which is the
medically most important toxin from the notorious black mamba (Dendroaspis polylepis), was employed.
Following immunization of a rabbit, a specific polyclonal antibody response was confirmed by ELISA and
immunodiffusion. Subsequently, these antibodies were investigated by high-density peptide microarray
to reveal linear elements of recognized epitopes across 742 3FTxs and 10 dendrotoxins. This exploratory
study demonstrates in a single immunized animal that cross-reactivity between toxins of high similarity
may be difficult to obtain when immunizing with a single 3FTx containing venom fraction. Additionally,
this study explored the influence of employing different lengths of peptides in high-density peptide
microarray experiments for identification of toxin epitopes. Using 8-mer, 12-mer, and 15-mer peptides, a
single linear epitope element was identified in SN-1 with high precision.