Molek Peter, Bratkovič Tomaž
Acta Chim Slov. 2016 Dec;63(4):914-919. doi: 10.17344/acsi.2016.2458.
Detailed knowledge of antigenic determinants is crucial when characterizing therapeutic and diagnostic antibodies, assessing vaccine effectiveness and developing epitope-based vaccines. Most epitope mapping approaches are labor intensive and costly. In this study, we evaluated panning of phage-displayed random peptide libraries against antibodies as a tool for cognate epitope identification. We used six antibodies directed to three model protein antigens as targets to show that the approach is applicable to both mono- and polyclonal antibodies. The technique is well-suited especially for identification of linear epitopes. Mapping of conformational epitopes is more challenging, tends to be more subjective and requires use of computational tools. Nevertheless, when combined with functional data such as structure-activity relationship of antigen muteins, one can make reliable conformational epitope predictions based on phage display experiment data. As the described approach is fast and relatively inexpensive, we suggest it is employed early in antibody characterization and later validated by complementary methods.
在表征治疗性和诊断性抗体、评估疫苗效力以及开发基于表位的疫苗时，对抗原决定簇的详细了解至关重要。大多数表位作图方法都需要大量人力且成本高昂。在本研究中，我们评估了利用噬菌体展示随机肽库筛选抗体作为识别同源表位的一种工具。我们使用针对三种模型蛋白抗原的六种抗体作为靶标，以表明该方法适用于单克隆抗体和多克隆抗体。该技术特别适合于线性表位的鉴定。构象表位的作图更具挑战性，往往更主观，并且需要使用计算工具。然而，当与诸如抗原突变体的构效关系等功能数据相结合时，就可以基于噬菌体展示实验数据做出可靠的构象表位预测。由于所描述的方法快速且相对便宜，我们建议在抗体表征的早期采用该方法，随后通过互补方法进行验证。
