Department of Bioscience and Biotechnology, University of Camerino, Camerino (MC), Italy.
PLoS One. 2013;8(3):e58358. doi: 10.1371/journal.pone.0058358. Epub 2013 Mar 28.
We developed a new phage-display based approach, the Large Fragment Phage Display (LFPD), that can be used for mapping conformational epitopes on target molecules of immunological interest. LFPD uses a simplified and more effective phage-display approach in which only a limited set of larger fragments (about 100 aa in length) are expressed on the phage surface. Using the human HER2 oncoprotein as a target, we identified novel B-cell conformational epitopes. The same homologous epitopes were also detected in rat HER2 and all corresponded to the epitopes predicted by computational analysis (PEPITO software), showing that LFPD gives reproducible and accurate results. Interestingly, these newly identified HER2 epitopes seem to be crucial for an effective immune response against HER2-overexpressing breast cancers and might help discriminating between metastatic breast cancer and early breast cancer patients. Overall, the results obtained in this study demonstrated the utility of LFPD and its potential application to the detection of conformational epitopes on many other molecules of interest, as well as, the development of new and potentially more effective B-cell conformational epitopes based vaccines.
我们开发了一种新的噬菌体展示技术，即大片段噬菌体展示（LFPD），可用于绘制免疫相关靶分子的构象表位。LFPD 采用简化且更有效的噬菌体展示方法，仅在噬菌体表面表达有限数量的较大片段（约 100 个氨基酸长）。我们用人源 HER2 癌蛋白作为靶标，鉴定了新的 B 细胞构象表位。在大鼠 HER2 中也检测到了相同的同源表位，并且都与计算分析（PEPITO 软件）预测的表位相对应，表明 LFPD 可获得可重复且准确的结果。有趣的是，这些新鉴定的 HER2 表位似乎对针对 HER2 过表达乳腺癌的有效免疫反应至关重要，并且可能有助于区分转移性乳腺癌和早期乳腺癌患者。总的来说，本研究的结果证明了 LFPD 的实用性及其在检测许多其他感兴趣的分子的构象表位以及基于新的潜在更有效的 B 细胞构象表位疫苗方面的应用潜力。
