Cardoso R, Homsi-Brandeburgo M I, Rodrigues V M, Santos W B, Souza G L R, Prudencio C R, Siquieroli A C S, Goulart L R
Institute of Genetics and Biochemistry, Federal University of Uberlândia, Minas Gerais, Brazil.
Toxicon. 2009 Feb;53(2):254-61. doi: 10.1016/j.toxicon.2008.11.009. Epub 2008 Dec 3.
Peptides derived from a phage display library may mimic essential features of epitopes (mimotopes), including their immunogenicity. A recombinant peptide library of 12 amino acids displayed on the phage capsid was used to obtain peptides that mimic epitopes of antigens that are reactive to specific polyclonal antibodies anti-neuwiedase (NEU), a toxin from Bothrops neuwiedi snake venom. These polyclonal antibodies are protective against NEU activity and were used as target for the peptide library biopannings, resulting in the selection of 80 peptides. Antibody-binding epitopes were obtained by sequence alignment with the primary and tertiary structures of the NEU protein. Antigenicity and specificity of the mimotopes mixture were confirmed by dot blot, immuno dot blot, plaque reduction and Western blot assays. Their immunogenicity was demonstrated by immunization of BALB/c mice and ELISA tests. The NEU toxin is an important antigen that has many common structural regions to several toxic venom metalloproteinases, in which two epitope regions have been detected. The two mapped epitopes were found in primary sequences of several snake venom toxins, thus demonstrating the potential application of these NEU mimotopes as possible antigen components that are toxicity free.
从噬菌体展示文库衍生的肽可能模拟表位的基本特征（模拟表位），包括它们的免疫原性。利用展示在噬菌体衣壳上的12个氨基酸的重组肽文库来获得模拟对巴西矛头蝮蛇毒中的一种毒素——新维德酶（NEU）有反应的抗原表位的肽。这些多克隆抗体对NEU活性具有保护作用，并被用作肽文库淘选的靶标，从而筛选出80种肽。通过与NEU蛋白的一级和三级结构进行序列比对获得抗体结合表位。通过斑点印迹、免疫斑点印迹、蚀斑减少和蛋白质印迹分析证实了模拟表位混合物的抗原性和特异性。通过对BALB/c小鼠进行免疫和ELISA试验证明了它们的免疫原性。NEU毒素是一种重要的抗原，与几种有毒毒液金属蛋白酶有许多共同的结构区域，其中已检测到两个表位区域。在几种蛇毒毒素的一级序列中发现了这两个定位的表位，因此证明了这些NEU模拟表位作为无毒的可能抗原成分的潜在应用。
