Durans Andressa da M, Napoleão-Pêgo Paloma, Reis Flavia C G, Dias Evandro R, Machado Luciana E S F, Lechuga Guilherme C, Junqueira Angela C V, De-Simone Salvatore G, Provance David W
Center for Technological Development in Health, National Institute of Science and Technology for Innovation in Neglected Population Diseases, Oswaldo Cruz Foundation, Rio de Janeiro 21040-900, Brazil.
Epidemiology and Molecular Systematics Laboratory, Oswaldo Cruz Institute, Oswaldo Cruz Foundation, Rio de Janeiro 21040-900, Brazil.
Vaccines (Basel). 2024 Sep 8;12(9):1029. doi: 10.3390/vaccines12091029.
Serological tests are critical tools in the fight against infectious disease. They detect antibodies produced during an adaptive immune response against a pathogen with an immunological reagent, whose antibody binding characteristics define the specificity and sensitivity of the assay. While pathogen proteins have conveniently served as reagents, their performance is limited by the natural grouping of specific and non-specific antibody binding sites, epitopes. An attractive solution is to build synthetic proteins that only contains pathogen-specific epitopes, which could theoretically reach 100% specificity. However, the genesis of de novo proteins remains a challenge. To address the uncertainty of producing a synthetic protein, we have repurposed the beta barrel of fluorescent proteins into a receptacle that can receive several epitope sequences without compromising its ability to be expressed. Here, two versions of a multiepitope protein were built using the receptacle that differ by their grouping of epitopes specific to the parasite , the causative agent for Chagas disease. An evaluation of their performance as the capture reagent in ELISAs showed near-complete agreement with recommended diagnostic protocols. The results suggest that a single assay could be developed for the diagnosis of Chagas disease and that this approach could be applied to other diseases.
血清学检测是对抗传染病的关键工具。它们用免疫试剂检测适应性免疫反应针对病原体产生的抗体，该免疫试剂的抗体结合特性决定了检测的特异性和灵敏度。虽然病原体蛋白一直方便地用作试剂，但其性能受到特异性和非特异性抗体结合位点（表位）自然分组的限制。一个有吸引力的解决方案是构建只包含病原体特异性表位的合成蛋白，理论上其特异性可达100%。然而，从头合成蛋白质仍然是一个挑战。为了解决合成蛋白质生产的不确定性，我们将荧光蛋白的β桶重新用作一个容器，该容器可以容纳多个表位序列而不影响其表达能力。在这里，使用该容器构建了两种多表位蛋白版本，它们在针对恰加斯病病原体（寄生虫）的表位分组上有所不同。对它们作为酶联免疫吸附测定（ELISA）中捕获试剂的性能评估表明，与推荐的诊断方案几乎完全一致。结果表明，可以开发一种单一检测方法用于恰加斯病的诊断，并且这种方法可以应用于其他疾病。
