Rojas Gertrudis, Tundidor Yaima, Infante Yanelys Cabrera
a Systems Biology Department ; Center of Molecular Immunology ; La Habana , Cuba.
MAbs. 2014;6(6):1368-76. doi: 10.4161/mabs.36144.
Antibody engineering must be accompanied by mapping strategies focused on identifying the epitope recognized by each antibody to define its unique functional identity. High throughput fine specificity determination remains technically challenging. We review recent experiences aimed at revisiting the oldest and most extended display technology to develop a robust epitope mapping platform, based on the ability to manipulate target-derived molecules (ranging from the whole native antigen to antigen domains and smaller fragments) on filamentous phages. Single, multiple and combinatorial mutagenesis allowed comprehensive scanning of phage-displayed antigen surface that resulted in the identification of clusters of residues contributing to epitope formation. Functional pictures of the epitope(s) were thus delineated in the natural context. Successful mapping of antibodies against interleukin-2, epidermal growth factor and its receptor, and vascular endothelial growth factor showed the versatility of these procedures, which combine the accuracy of site-directed mutagenesis with the high throughput potential of phage display.
抗体工程必须辅以定位策略，重点是识别每种抗体所识别的表位，以确定其独特的功能特性。高通量精细特异性测定在技术上仍然具有挑战性。我们回顾了最近的一些经验，旨在重新审视最古老、应用最广泛的展示技术，以基于在丝状噬菌体上操纵靶标衍生分子（从完整的天然抗原到抗原结构域和更小的片段）的能力，开发一个强大的表位定位平台。单突变、多突变和组合突变允许对噬菌体展示的抗原表面进行全面扫描，从而识别出有助于表位形成的残基簇。因此，在自然环境中描绘出了表位的功能图谱。针对白细胞介素-2、表皮生长因子及其受体以及血管内皮生长因子的抗体的成功定位显示了这些方法的通用性，这些方法将定点突变的准确性与噬菌体展示的高通量潜力相结合。
