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Multifunctional Dendrimer-Templated Antibody Presentation on Biosensor Surfaces for Improved Biomarker Detection

机译：多功能树状大分子模板抗体在生物传感器表面上的呈现，以改善生物标志物的检测。

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Dendrimers, with their well-defined globular shape and high density of functional groups, are ideal nanoscale materials for templating sensor surfaces. This work exploits dendrimers as a versatile platform for capturing biomarkers with improved sensitivity and specificity. The synthesis, characterization, fabrication, and functional validation of the dendrimer-based assay platform are described. Bifunctional hydroxyl/thiol-functionalized G4-polyamidoamine (PAMAM) dendrimer is synthesized and immobilized on the polyethylene-glycol (PEG)-functionalized assay plate by coupling PEG-maleimide and dendrimer thiol groups. Simultaneously, part of the dendrimer thiol groups are converted to hydrazide functionalities. The resulting dendrimer-modified surface is coupled to the capture antibody in the Fc region of the oxidized antibody. This preserves the orientation flexibility of the antigen binding region (Fv) of the antibody. To validate the approach, the fabricated plates are further used as a solid phase for developing a sandwich-type enzyme-linked immunosorbent assay (ELISA) to detect IL-6 and IL-1β, important biomarkers for early stages of chorioamnionitis. The dendrimer-modified plate provides assays with significantly enhanced sensitivity, lower nonspecific adsorption, and a detection limit of 0.13 pg mL~(-1) for IL-6 luminol detection and 1.15 pg mL~(-1) for IL-1β TMB detection, which are significantly better than those for the traditional ELISA. The assays were validated in human serum samples from a normal (nonpregnant) woman and pregnant women with pyelonephritis. The specificity and the improved sensitivity of the dendrimer-based capture strategy could have significant implications for the detection of a wide range of cytokines and biomarkers since the capture strategy could be applied to multiplex microbead assays, conductometric immunosensors, and field-effect biosensors.
机译：树枝状聚合物具有定义明确的球形形状和高密度的官能团，是用于模板化传感器表面的理想纳米级材料。这项工作利用树状聚合物作为捕获生物标志物的通用平台，具有更高的灵敏度和特异性。描述了基于树枝状聚合物的测定平台的合成，表征，制备和功能验证。合成双功能羟基/硫醇官能化的G4-聚酰胺型（PAMAM）树状聚合物，并通过偶联PEG-马来酰亚胺和树状聚合物硫醇基团将其固定在聚乙二醇（PEG）功能化的测定板上。同时，部分树状聚合物硫醇基团转化为酰肼官能团。所得的树状聚合物修饰的表面在氧化抗体的Fc区域中与捕获抗体偶联。这保留了抗体的抗原结合区（Fv）的定向柔性。为了验证该方法，将制成的板进一步用作开发夹心型酶联免疫吸附测定（ELISA）的固相，以检测IL-6和IL-1β（绒毛膜羊膜炎早期的重要生物标志物）。树枝状大分子修饰板提供了显着提高的灵敏度，更低的非特异性吸附，IL-6鲁米诺检测限为0.13 pg mL〜（-1）和IL-1βTMB检测限为1.15 pg mL〜（-1）的检测方法，这比传统ELISA的要好得多。该测定法在来自正常（未怀孕）妇女和患有肾盂肾炎的孕妇的人血清样品中得到验证。基于树状大分子的捕获策略的特异性和更高的灵敏度可能对多种细胞因子和生物标志物的检测具有重要意义，因为捕获策略可应用于多重微珠测定，电导免疫传感器和场效应生物传感器。

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  • 来源
    《Advanced Functional Materials》 |2010年第3期|409-421|共13页
  • 作者

    Hye Jung Han; Rangaramanujam M. Kannan; Sunxi Wang; Guangzhao Mao; Juan Pedro Kusanovic; Roberto Romero;

  • 作者单位

    Department of Chemical Engineering and Materials Science Wayne State University Detroit, MI 48202 (USA) Eunice Kennedy Shriver National Institute of Child Health and Human Development NICHD, NIH, DHHS Detroit, MI 48201 (USA);

    Department of Chemical Engineering and Materials Science Wayne State University Detroit, MI 48202 (USA) Eunice Kennedy Shriver National Institute of Child Health and Human Development NICHD, NIH, DHHS Detroit, MI 48201 (USA);

    Department of Chemical Engineering and Materials Science Wayne State University Detroit, MI 48202 (USA);

    Department of Chemical Engineering and Materials Science Wayne State University Detroit, MI 48202 (USA);

    Perinatology Research Branch NICHD/NIH/DHHS Department of Obstetrics and Gynecology Wayne State University, Hutzel Women's Hospital Detroit, MI 48201 (USA);

    Eunice Kennedy Shriver National Institute of Child Health and Human Development NICHD, NIH, DHHS Detroit, MI 48201 (USA);

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