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首页> 外文期刊>RSC Advances >Development of a nanobody tagged with streptavidin-binding peptide and its application in a Luminex fluoroimmunoassay for alpha fetal protein in serum
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Development of a nanobody tagged with streptavidin-binding peptide and its application in a Luminex fluoroimmunoassay for alpha fetal protein in serum

机译：用链霉抗生物素蛋白结合肽标记的纳米曲面的研制及其在血清中α胎儿蛋白的Luminex FluoroImmunoassay中的应用

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Sensitive and accurate detection of disease-related biomarkers can promote the early screening and diagnosis of cancers for improving the prognosis and survival of patients. Herein alpha fetal protein (AFP) was selected as the model macromolecule antigen and we developed AFP-specific alpaca nanobodies (Nbs) from an immunized phage-displayed Nb library. Then Nbs tagged with streptavidin-binding peptide (Nb-SBP) were constructed and used to develop an Nb-SBP-mediated fluoroimmunoassay based on the Luminex-200 system (NS-LFIA). Based on the optimal experimental conditions, the NS-LFIA has a limit of detection of 0.237 ng mL(-1)with a linear detection range of 0.49-125 ng mL(-1). The average recovery rate and relative standard derivation were in the range of 98.2-110% and 2.8-13.8%, respectively. The NS-LFIA is highly selective for AFP and ignorable cross-reaction was observed with the other biomarkers. The content of AFP in clinical serum samples was determined by both the developed NS-LFIA and the Roche E601 automatic chemiluminescence immunoassay analyzer and a good correlation was obtained between the two methods (R-2= 0.9894). Moreover, the Nb-SBP can significantly improve the homogeneity of the fluorescent signals tested by the Luminex-200 system compared with the biotinylated conventional monoclonal antibodies, which could reduce the magnetic microsphere consumption and test cost by decreasing the repetitions of each sample. Thus the results demonstrated that the Nb-SBP was a very promising immunological diagnostic reagent and indicated the applicability and reliability of the NS-LFIA for sensitive detection of AFP and other disease-related biomarkers.
机译：敏感和准确的疾病相关生物标志物检测可以促进早期筛查和诊断癌症，以改善患者的预后和存活。在此选择α胎儿蛋白（AFP）作为模型大分子抗原，并且我们从免疫噬菌体显示的Nb文库中开发了AFP特异性的羊驼纳米菌（NBS）。然后构建标记用链霉蛋白结合肽（NB-SBP）的NBS，并用于基于Luminex-200系统（NS-LFIA）进行Nb-Sbp介导的氟莫莫纳斯。基于最佳实验条件，NS-LFIA的检测限为0.237ng ml（-1），线性检测范围为0.49-125 ng ml（-1）。平均回收率和相对标准衍生分别为98.2-110％和2.8-13.8％。 NS-LFIa对AFP的高度选择性，并且用其他生物标志物观察到无知的交叉反应。临床血清样品中AFP的含量由发育的NS-LFIA和Roche E601自动化学发光免疫测定分析仪测定，并且在两种方法之间获得了良好的相关性（R-2 = 0.9894）。此外，与生物素化的常规单克隆抗体相比，Nb-Sbp可以显着改善Luminex-200系统测试的荧光信号的均匀性，这可以通过降低每个样品的重复来降低磁体微球消耗和测试成本。因此，结果表明，Nb-SBP是一种非常有前景的免疫诊断试剂，并表明NS-LFIA的适用性和可靠性用于AFP和其他与其他疾病相关的生物标志物的敏感性检测。

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  • 来源
    《RSC Advances》 |2020年第40期|共8页
  • 作者

    Chen Qi; Sun Danyang; Pei Hua; Su Benchao; Bao Kunlu; Cao Hongmei; Zhang Chenghui; Hammock Bruce D.; Liu Xing;

  • 作者单位

    Hainan Univ Coll Food Sci &

    Engn Haikou 570228 Hainan Peoples R China;

    Hainan Univ Coll Food Sci &

    Engn Haikou 570228 Hainan Peoples R China;

    Hainan Med Univ Dept Clin Lab Affiliated Hosp 2 Haikou 570311 Hainan Peoples R China;

    Hainan Univ Coll Food Sci &

    Engn Haikou 570228 Hainan Peoples R China;

    Hainan Univ Coll Food Sci &

    Engn Haikou 570228 Hainan Peoples R China;

    Hainan Univ Coll Food Sci &

    Engn Haikou 570228 Hainan Peoples R China;

    Hainan Univ Coll Food Sci &

    Engn Haikou 570228 Hainan Peoples R China;

    Univ Calif Davis Dept Entomol &

    Nematol Davis CA 95616 USA;

    Hainan Univ Coll Food Sci &

    Engn Haikou 570228 Hainan Peoples R China;

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