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首页> 外文期刊>Biomaterials Science >Conjugation of antibody with temperature-responsive polymer via in situ click reaction to enable biomarker enrichment for increased diagnostic sensitivity
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Conjugation of antibody with temperature-responsive polymer via in situ click reaction to enable biomarker enrichment for increased diagnostic sensitivity

机译：通过原位点击反应将抗体与温度响应聚合物的缀合，以使生物标志物富集以增加诊断敏感性

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摘要

Early diagnosis of infectious diseases is one of the current prevalent challenges, especially in low and limited resource settings where simple, fast, portable, cheap, and sensitive diagnostic approaches are needed. Lateral flow immunoassay (LFIA) is a common, rapid screening assay. However, the low assay sensitivity limits the utility of LFIA for specimens with low pathogenic loads (early infection stages). Antibodies conjugated with stimulus-responsive polymers have been previously utilized to improve assay sensitivity for detection of biomarkers at low concentrations. However, the loss of antibody affinity after polymer conjugation remains a significant challenge. In this study, we developed poly(N-isopropylacrylamide-co-N-(2-hydroxyisopropyl)acrylamide-co-strained alkyne-isopropylacrylamide), a novel polymer for biomarker enrichment, by polymer conjugation after antibody-antigen recognition. We employed and promoted the click chemistry in situ, to facilitate highly specific conjugation between novel temperature-responsive polymers and antibody-antigen complexes. This method could suppress the decrease in the binding constant associated with polymer conjugation (>20-fold). The conjugation was successfully demonstrated in body fluids such as urine and saliva. We achieved >5-fold antigen enrichment via thermal precipitation by conjugating polymers to the antibodies after antigen recognition. Concentrated biomarkers resulted in improved LFIA detection. This approach can potentially be utilized to improve diagnostic tests for infectious diseases in low and limited resource settings.
机译：早期诊断传染病是当前普遍的挑战之一，特别是在低价和有限的资源环境中，需要简单，快速，便携，便宜和敏感的诊断方法。外侧流动免疫测定（LFIa）是常见的快速筛选测定。然而，低测定敏感性限制了LFIa对具有低致病载荷（早期感染阶段）的试样的效用。先前已经利用缀合与刺激响应性聚合物的抗体以改善测定敏感性，以在低浓度下检测生物标志物。然而，聚合物缀合后的抗体亲和力的丧失仍然是重大挑战。在这项研究中，我们通过聚合物缀合在抗体 - 抗原识别后，开发了聚（N-异丙基丙烯酰胺-CO-（2-羟基异丙基）丙烯酰胺 - 共克炔类丙烯酰胺，用于生物标志物富集的新型聚合物。我们使用并促进了咔哒化学原位，以促进新型温度响应聚合物和抗体 - 抗原复合物之间的高度特异性缀合。该方法可以抑制与聚合物共轭（> 20倍）相关的结合常数的降低。在体液中成功证明了缀合物，例如尿液和唾液。通过将聚合物缀合在抗原识别后通过热沉淀来实现> 5倍抗原富集。浓缩的生物标志物导致LFIa检测改善。该方法可能用于改善低和有限的资源环境中传染病的诊断测试。

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  • 来源
    《Biomaterials Science》 |2021年第14期|共11页
  • 作者

    Hironaka Keita; Yoshihara Erika; Nabil Ahmed; Lai James J.; Kikuchi Akihiko; Ebara Mitsuhiro;

  • 作者单位

    Natl Inst Mat Sci NIMS Res Ctr Funct Mat RCFM Tsukuba Ibaraki 3050044 Japan;

    Natl Inst Mat Sci NIMS Res Ctr Funct Mat RCFM Tsukuba Ibaraki 3050044 Japan;

    Natl Inst Mat Sci NIMS Res Ctr Funct Mat RCFM Tsukuba Ibaraki 3050044 Japan;

    Univ Washington Ctr Bioengn Seattle WA 98195 USA;

    Tokyo Univ Sci Grad Sch Ind Sci &

    Technol Dept Mat Sci &

    Technol 6-3-1 Niijuku Tokyo 1258585 Japan;

    Natl Inst Mat Sci NIMS Res Ctr Funct Mat RCFM Tsukuba Ibaraki 3050044 Japan;

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  • 正文语种 eng
  • 中图分类 分子生物学;
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