Instituto de Biomedicina de Valencia, CSIC, Valencia, Spain.
Toxicon. 2012 Sep 15;60(4):688-99. doi: 10.1016/j.toxicon.2012.04.342. Epub 2012 Apr 26.
A second generation antivenomics protocol, based on affinity chromatography, was compared with a previously (first generation) immunodepletion protocol using as a proof of principle the pan-African EchiTAb-Plus-ICP(®) IgG antivenom and the venoms of Echis ocellatus, Bitis arietans, and African spitting cobras. The antivenom showed qualitatively similar immunoreactivity patterns using either antivenomic approach. Quantitative departures were noticed between both methods, which may be ascribed to differences in calculating the relative amounts of the non-recognized venom proteins. The smoother baseline in chromatograms of the affinity column allowed better resolution and more accurate quantification of the antivenomic outcome than the original immunodepletion protocol. Our results indicate that both methods can be used interchangeably to investigate the in vitro immunoreactivity of antivenoms. However, advantages of the second generation antivenomics are the possibility of analyzing F(ab')(2) antivenoms and the reusability of the affinity columns. These features contribute to the generalization, economy and reproducibility of the method.
第二代亲和层析型抗蛇毒血清组学方案与之前的（第一代）免疫沉淀方案进行了比较，研究原理采用泛非 EchiTAb-Plus-ICP(®) IgG 抗蛇毒血清和 Echis ocellatus、Bitis arietans 和非洲喷毒眼镜蛇的毒液。使用抗蛇毒血清组学方法时，抗蛇毒血清表现出定性上相似的免疫反应模式。两种方法之间存在定量差异，这可能归因于计算未识别的毒液蛋白相对量的差异。亲和柱色谱图中的基线更平滑，与原始免疫沉淀方案相比，它可以更好地分离和更准确地定量抗蛇毒血清组学结果。我们的结果表明，这两种方法可以互换使用，以研究抗蛇毒血清的体外免疫反应。然而，第二代抗蛇毒血清组学的优势在于可以分析 F(ab')(2) 抗蛇毒血清和重复使用亲和柱。这些特点有助于方法的推广、经济化和重现性。
