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用于高性能 ELISA 的功能轻量化聚苯乙烯@聚多巴胺纳米颗粒。

Functional lightweight polystyrene@polydopamine nanoparticle for high-performance ELISA.

机构信息

Hunan Provincial Key Laboratory of Micro & Nano Materials Interface Science, College of Chemistry and Chemical Engineering, Central South University, Changsha, Hunan Province, 410083, PR China.

The Department of Dermatology, Xiangya Hospital, Central South University, Changsha, 410008, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, 410008, China.

出版信息

Talanta. 2023 Jan 15;252:123871. doi: 10.1016/j.talanta.2022.123871. Epub 2022 Aug 25.

Abstract

Nanoparticles are usually used as carrier to load more antibody and enzyme to improve the sensitivity of enzyme-linked immunosorbent assay (ELISA). However, limited by high density and complicated modification procedure, the traditional nanoparticles such as Au nanoparticles (AuNPs) usually induce large background signal and poor reproducibility in ELISA. In this work, functional lightweigh nanoparticle polystyrene@polydopamine (PS@PDA) was prepared and induced as the carrier of detection antibody and horseradish peroxidase (HRP) to form PS@PDA@Ab/HRP biojungates. The appropriate density (close to water) and good hydropilicity ensure the good dispersion of PS@PDA@Ab/HRP in solution, preventing the physical sedimention and decreasing the background signal even though the bioconjugate's size is close to 200 nm. The large surface area and abundant active group from PDA facilitate the loading of detection antibody and HRP, improving the loading efficiency and stability of biojungates. Based on it, taking interleukin-17A (IL-17A, a biomarker of psoriasis) as the detection target, we developed a PS@PDA-based sandwich ELISA, achieving a sensitive dynamic range from 0.3 to 80 pg/mL and a detection limit of 0.2 pg/mL. Furthermore, the contents of IL-17A were assayed successfully in 10-fold diluted serum samples from psoriasis patients. Compared with those commercial or AuNP-based ELISA, our PS@PDA-based ELISA method exhibits higher sensitivity, lower background interference, and higher stability, which will significantly improve the application of ELISA in the low-abundant biomolecule assays.

摘要

纳米粒子通常被用作载体来负载更多的抗体和酶,以提高酶联免疫吸附测定(ELISA)的灵敏度。然而,受限于高密度和复杂的修饰过程,传统的纳米粒子,如金纳米粒子(AuNPs),通常会在 ELISA 中引起较大的背景信号和较差的重现性。在这项工作中,制备了功能化的轻质纳米粒子聚苯乙烯@聚多巴胺(PS@PDA),并将其作为检测抗体和辣根过氧化物酶(HRP)的载体,形成 PS@PDA@Ab/HRP 生物缀合物。适当的密度(接近水)和亲水性确保了 PS@PDA@Ab/HRP 在溶液中的良好分散性,防止了物理沉降,即使生物缀合物的大小接近 200nm,也能降低背景信号。PDA 丰富的表面活性基团和大量的活性基团有利于检测抗体和 HRP 的负载,提高了生物缀合物的负载效率和稳定性。基于此,以白细胞介素-17A(IL-17A,银屑病的生物标志物)为检测目标,我们开发了一种基于 PS@PDA 的夹心 ELISA,实现了 0.3 至 80pg/mL 的灵敏动态范围和 0.2pg/mL 的检测限。此外,还成功地测定了 10 倍稀释的银屑病患者血清样本中的 IL-17A 含量。与那些商业 ELISA 或基于 AuNP 的 ELISA 相比,我们基于 PS@PDA 的 ELISA 方法具有更高的灵敏度、更低的背景干扰和更高的稳定性,这将显著提高 ELISA 在低丰度生物分子检测中的应用。

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