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整合抗体与催化金属有机骨架的比色免疫分析

Integrated Antibody with Catalytic Metal-Organic Framework for Colorimetric Immunoassay.

机构信息

Key Laboratory of Chemical Biology of Jiangxi Province, College of Chemistry and Chemical Engineering , Jiangxi Normal University , Nanchang 330022 , P. R. China.

出版信息

ACS Appl Mater Interfaces. 2018 Aug 1;10(30):25113-25120. doi: 10.1021/acsami.8b07225. Epub 2018 Jul 20.

Abstract

Enzyme-linked immunosorbent assay has been widely used as a gold standard in biomedical field, but some inevitable drawbacks still exist in its practical applications, especially the laborious preparation of enzyme-antibody conjugates by a covalent linkage. In this work, we proposed a new strategy to prepare enzyme-antibody conjugate by integrating antibody with catalytic metal-organic framework (MOF) to form dual-functional MOF/antibody composite. As models, rabbit antimouse immunoglobulin G antibody (RIgG) and Cu-MOF with peroxidase-like activity were used to fabricate RIgG@Cu-MOF composite for colorimetric immunoassay. It was found that Cu-MOF as a host not only has no influence on the original capture ability of RIgG to its corresponding antigen (mIgG), but also can shield RIgG against long-term storage, high temperature, and biological degradation. More importantly, upon the formation of sandwiched immunocomplex between RIgG@Cu-MOF and capture antibody, Cu-MOF can serve as a signal amplification unit to perform colorimetric immunoassay. The detection limit of RIgG@Cu-MOF toward mIgG was obtained at 0.34 ng/mL, which is 3-fold lower than that of horseradish peroxidase labeled RIgG. Furthermore, the successful determination of mIgG in serum sample demonstrates the applicability of RIgG@Cu-MOF in detecting real sample. Therefore, it is highly anticipated that this study can offer a new way to prepare enzyme-antibody conjugates, facilitating the exploration of MOF composites in biomedical field.

摘要

酶联免疫吸附测定法已被广泛应用于生物医学领域,被视为金标准,但在实际应用中仍存在一些不可避免的缺陷,特别是酶-抗体偶联物的制备需要通过共价键连接,这一过程较为繁琐。在这项工作中,我们提出了一种新策略,通过将抗体与具有类过氧化物酶活性的催化金属-有机骨架(MOF)整合来制备酶-抗体偶联物,从而形成双功能的 MOF/抗体复合材料。以兔抗鼠免疫球蛋白 G 抗体(RIgG)和具有过氧化物酶样活性的 Cu-MOF 为模型,制备了用于比色免疫分析的 RIgG@Cu-MOF 复合材料。研究发现,Cu-MOF 作为主体,不仅对 RIgG 与其相应抗原(mIgG)的原始捕获能力没有影响,而且还可以保护 RIgG 免受长期储存、高温和生物降解的影响。更为重要的是,在 RIgG@Cu-MOF 和捕获抗体形成夹心免疫复合物后,Cu-MOF 可以作为信号放大单元进行比色免疫分析。RIgG@Cu-MOF 对 mIgG 的检测限达到 0.34 ng/mL,比辣根过氧化物酶标记的 RIgG 低 3 倍。此外,成功地测定了血清样品中的 mIgG,证明了 RIgG@Cu-MOF 在检测实际样品中的适用性。因此,这项研究有望为制备酶-抗体偶联物提供一种新方法,促进 MOF 复合材料在生物医学领域的探索。

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