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一种用于检测抗弓形虫免疫球蛋白的新型基于肽的传感器平台。

A novel peptide-based sensor platform for detection of anti-Toxoplasma gondii immunoglobulins.

机构信息

Institute of Biotechnology, Federal University of Uberlândia, Uberlândia, Brazil.

Institute of Biomedical Sciences, Federal University of Uberlândia, Uberlândia, Brazil.

出版信息

J Pharm Biomed Anal. 2019 Oct 25;175:112778. doi: 10.1016/j.jpba.2019.112778. Epub 2019 Jul 17.

Abstract

Toxoplasma gondii is an intracellular protozoan parasite responsible for toxoplasmosis, which affects humans and animals. Serologic detection of anti-T. gondii immunoglobulins plays a crucial role in the clinical diagnosis of toxoplasmosis. In this work, a novel electrochemical immunosensor for detecting anti-T. gondii immunoglobulins is reported, based on immobilization of an in silico predicted peptide (PepB3), obtained from membrane protein of T. gondii, on the graphite electrode modified with poly(3-hydroxybenzoic acid). Indirect ELISA confirmed infection and binding specificity of peptide PepB3. Molecular modelling and simulations show this peptide binds to the T. gondii human Fab antibody in the surface antigen 1 (SAG1) binding site, remaining a stable complex during the molecular dynamic simulations, especially by hydrogen bonds and hydrophobic interactions. This electrochemical immunosensor was able to discriminate different periods of infection, using infected mouse serum samples, showing selectivity and discriminating infected and uninfected mouse serum.

摘要

刚地弓形虫是一种细胞内的原生动物寄生虫,可引起弓形体病,影响人类和动物。抗刚地弓形虫免疫球蛋白的血清学检测在弓形体病的临床诊断中起着至关重要的作用。在这项工作中,报道了一种基于硅预测肽(PepB3)固定在聚(3-羟基苯甲酸)修饰的石墨电极上的新型电化学免疫传感器,用于检测抗刚地弓形虫免疫球蛋白。间接 ELISA 证实了肽 PepB3 的感染和结合特异性。分子建模和模拟表明,这种肽结合到刚地弓形虫人类 Fab 抗体在表面抗原 1(SAG1)结合位点,在分子动力学模拟过程中保持稳定的复合物,特别是通过氢键和疏水相互作用。该电化学免疫传感器能够使用感染的小鼠血清样本区分不同的感染期,具有选择性,并区分感染和未感染的小鼠血清。

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