Fogaça Rafaela L, Capelli-Peixoto Janaína, Yamanaka Isabel B, de Almeida Rodrigo P M, Muzzi João Carlos D, Borges Mariangela, Costa Alvimar J, Chávez-Olortegui Carlos, Thomaz-Soccol Vanete, Alvarenga Larissa M, de Moura Juliana
Departamento de Patologia Básica, UFPR, Curitiba, PR, Brazil.
Appl Microbiol Biotechnol. 2014 Nov;98(21):8887-94. doi: 10.1007/s00253-014-5968-0. Epub 2014 Aug 2.
Bovine cysticercosis is detected during the routine post mortem examination of carcasses by visual inspection (knife and eye method). However, the sensitivity of this procedure is several times lower than immunoassays, even when it is performed by qualified professionals. In the present study, a new generation capture antigens were screened from a phage display peptide library using antibodies from Taenia saginata-infected animals. Eight phage clones were selected, and one, Tsag 3 (VHTSIRPRCQPRAITPR), produced similar results to the T. saginata metacestode crude antigen (TsCa) when used as a capture antigen in an ELISA. The phage-displayed peptides competed with TsCa for binding sites, reducing the reactivity by approximately 30 %. Alanine scanning indicated that proline, arginine, and serine are important residues for antibody binding. Tsag 1 (HFYQITWLPNTFPAR), the most frequent affinity-selected clone, and Tsag 6 (YRWPSTPSASRQATL) shared similarity with highly conserved proteins from the Taeniidae family with known immunogenicity. Due to their epitopic or mimotopic properties, these affinity-selected phages could contribute to the rational design of an ante mortem immunodiagnosis method for bovine cysticercosis, as well as an epitope-based vaccine to interrupt the taeniosis/cysticercosis complex.
牛囊尾蚴病是在屠体常规尸检过程中通过目视检查(刀检和眼检法)发现的。然而,即使由合格的专业人员进行操作,该方法的灵敏度也比免疫测定法低几倍。在本研究中,使用来自牛带绦虫感染动物的抗体从噬菌体展示肽库中筛选新一代捕获抗原。选择了8个噬菌体克隆,其中一个Tsag 3(VHTSIRPRCQPRAITPR)在ELISA中用作捕获抗原时,产生的结果与牛带绦虫囊尾蚴粗抗原(TsCa)相似。噬菌体展示的肽与TsCa竞争结合位点,使反应性降低约30%。丙氨酸扫描表明,脯氨酸、精氨酸和丝氨酸是抗体结合的重要残基。最常被亲和选择的克隆Tsag 1(HFYQITWLPNTFPAR)和Tsag 6(YRWPSTPSASRQATL)与具有已知免疫原性的带科家族高度保守蛋白具有相似性。由于它们的表位或模拟表位特性,这些亲和选择的噬菌体有助于合理设计牛囊尾蚴病生前免疫诊断方法以及基于表位的疫苗,以阻断绦虫病/囊尾蚴病复合体。
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