Canali Elena, Bolchi Angelo, Spagnoli Gloria, Seitz Hanna, Rubio Ivonne, Pertinhez Thelma A, Müller Martin, Ottonello Simone
1] Department of Life Sciences, Biochemistry and Molecular Biology Unit, University of Parma, Italy [2].
Department of Life Sciences, Biochemistry and Molecular Biology Unit, University of Parma, Italy.
Sci Rep. 2014 Apr 22;4:4729. doi: 10.1038/srep04729.
Escherichia coli thioredoxin has been previously exploited as a scaffold for the presentation/stabilization of peptide aptamers as well as to confer immunogenicity to peptide epitopes. Here we focused on other key features of thioredoxin that are of general interest for the production of safer and more effective peptide immunogens, such as a high thermal stability, lack of cross-reactivity and a low-cost of production. We identified thioredoxin from the archaebacterium Pyrococcus furiosus (PfTrx) as a novel scaffold meeting all the above criteria. PfTrx is a highly thermostable and protease-resistant scaffold with a strong (poly)peptide solubilisation capacity. Anti-PfTrx antibodies did not cross-react with mouse, nor human thioredoxin. Untagged PfTrx bearing a previously identified HPV16-L2 peptide epitope was obtained in a >90% pure form with a one-step thermal purification procedure and effectively elicited the production of neutralizing anti-HPV antibodies. We thus propose PfTrx as a superior, general-purpose scaffold for the construction of safe, stable, and low-cost peptide immunogens.
大肠杆菌硫氧还蛋白此前已被用作展示/稳定肽适配体以及赋予肽表位免疫原性的支架。在此，我们关注硫氧还蛋白的其他关键特性，这些特性对于生产更安全、更有效的肽免疫原具有普遍意义，例如高热稳定性、无交叉反应性以及低成本生产。我们从嗜热栖热菌（Pyrococcus furiosus，PfTrx）中鉴定出硫氧还蛋白作为满足上述所有标准的新型支架。PfTrx是一种高度耐热且抗蛋白酶的支架，具有很强的（多）肽溶解能力。抗PfTrx抗体与小鼠或人硫氧还蛋白均无交叉反应。携带先前鉴定出的HPV16 - L2肽表位的无标签PfTrx通过一步热纯化程序以>90%的纯度获得，并有效引发了中和性抗HPV抗体的产生。因此，我们提议将PfTrx作为构建安全、稳定且低成本肽免疫原的优质通用支架。
