Department of Pharmaceutical Biology, Faculty of Pharmacy, Aškerčeva 7, Ljubljana, Slovenia.
Molecules. 2011 Jan 19;16(1):790-817. doi: 10.3390/molecules16010790.
An increasing number of peptides with specific binding affinity to various protein and even non-protein targets are being discovered from phage display libraries. The power of this method lies in its ability to efficiently and rapidly identify ligands with a desired target property from a large population of phage clones displaying diverse surface peptides. However, the search for the needle in the haystack does not always end successfully. False positive results may appear. Thus instead of specific binders phage with no actual affinity toward the target are recovered due to their propagation advantages or binding to other components of the screening system, such as the solid phase, capturing reagents, contaminants in the target sample or blocking agents, rather than the target. Biopanning experiments on different targets performed in our laboratory revealed some previously identified and many new target-unrelated peptide sequences, which have already been frequently described and published, but not yet recognized as target-unrelated. Distinguishing true binders from false positives is an important step toward phage display selections of greater integrity. This article thoroughly reviews and discusses already identified and new target-unrelated peptides and suggests strategies to avoid their isolation.
越来越多的具有与各种蛋白质甚至非蛋白质靶标特异性结合亲和力的肽，正在从噬菌体展示文库中被发现。这种方法的威力在于，它能够从大量展示多样化表面肽的噬菌体克隆中，高效快速地识别具有所需靶标特性的配体。然而，大海捞针并不总能成功。可能会出现假阳性结果。因此，由于噬菌体的增殖优势或与筛选系统的其他成分（如固相、捕获试剂、靶样品中的污染物或阻断剂）结合，而不是与靶标结合，而不是特异性结合剂的噬菌体被回收。我们实验室在不同靶标上进行的生物淘选实验揭示了一些以前鉴定出的和许多新的与靶标无关的肽序列，这些序列已经被频繁描述和发表，但尚未被认为与靶标无关。将真正的结合物与假阳性区分开来，是进行噬菌体展示选择以获得更高完整性的重要步骤。本文全面回顾和讨论了已经鉴定出的和新的与靶标无关的肽，并提出了避免其分离的策略。
