Titus James K, Kay Matthew K, Glaser Cdr Jacob J
Naval Medical Research Unit San Antonio, 3650 Chambers Pass, Fort Sam Houston, Texas 78234, USA.
J Venom Res. 2017 Sep 28;8:19-24. eCollection 2017.
Snakebite envenomation is an important global health concern. The current standard treatment approach for snakebite envenomation relies on antibody-based antisera, which are expensive, not universally available, and can lead to adverse physiological effects. Phage display techniques offer a powerful tool for the selection of phage-expressed peptides, which can bind with high specificity and affinity towards venom components. In this research, the amino acid sequences of Phospholipase A (PLA) from multiple cottonmouth species were analyzed, and a consensus peptide synthesized. Three phage display libraries were panned against this consensus peptide, crosslinked to capillary tubes, followed by a modified surface panning procedure. This high throughput selection method identified four phage clones with anti-PLA activity against Western cottonmouth venom, and the amino acid sequences of the displayed peptides were identified. This is the first report identifying short peptide sequences capable of inhibiting PLA activity of Western cottonmouth venom , using a phage display technique. Additionally, this report utilizes synthetic panning targets, designed using venom proteomic data, to mimic epitope regions. M13 phages displaying circular 7-mer or linear 12-mer peptides with antivenom activity may offer a novel alternative to traditional antibody-based therapy.
蛇咬伤中毒是一个重要的全球健康问题。目前蛇咬伤中毒的标准治疗方法依赖于基于抗体的抗血清，这种抗血清价格昂贵，并非普遍可得，而且可能导致不良生理效应。噬菌体展示技术为筛选噬菌体表达的肽提供了一个强大工具，这些肽能够以高特异性和亲和力与毒液成分结合。在本研究中，分析了多种棉口蛇属物种的磷脂酶A（PLA）的氨基酸序列，并合成了一个共有肽。针对这个与毛细管交联的共有肽淘选了三个噬菌体展示文库，随后采用改良的表面淘选程序。这种高通量筛选方法鉴定出四个对西部棉口蛇毒液具有抗PLA活性的噬菌体克隆，并确定了所展示肽的氨基酸序列。这是首次使用噬菌体展示技术鉴定出能够抑制西部棉口蛇毒液PLA活性的短肽序列的报告。此外，本报告利用根据毒液蛋白质组学数据设计的合成淘选靶点来模拟表位区域。展示具有抗蛇毒活性的环状7肽或线性12肽的M13噬菌体可能为传统的基于抗体的治疗提供一种新的替代方法。
