A chemiluminescence assay for determination of lysozyme based on the use of magnetic alginate-aptamer composition and he
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ORIGINAL PAPER
A chemiluminescence assay for determination of lysozyme based on the use of magnetic alginate-aptamer composition and hemin@HKUST-1 Yanna Lin 1 & Yuanling Sun 1 & Yuxue Dai 1 & Xiaodong Zhu 1 & Hao Liu 1 & Rui Han 1 & Dandan Gao 1 & Chuannan Luo 1 & Xueying Wang 1 Received: 2 October 2019 / Accepted: 31 March 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract Lysozyme aptamer-functionalized magnetic alginate hydrogel was prepared for separation and enrichment of lysozyme. Luminol-labeled aptamer was used as a signal tag, and the signal tag was adsorbed on magnetic carboxylated carbon nanotubes based on the π-interaction. When lysozyme was added, the aptamer specifically binds to the lysozyme, causing the signal tag to detach from the magnetic carboxylated carbon nanotubes. When the aptamer/lysozyme complex bound to the complementary single strand of aptamer on the hemin@HKUST-1, lysozyme was released. The released lysozyme can be recombined with the signal tag adsorbed on the magnetic carboxylated carbon nanotube, allowing more signal tag to be dispersed into the solution. Determination of lysozyme was achieved by releasing the luminol-labeled aptamer to generate a chemiluminescence signal at a wavelength of 425 nm. It was proved by experiments that the synthesized hemin@HKUST-1 had a strong catalytic effect on the luminol-NaOH-H2O2 system. The chemiluminescence signal was increased nearly 100 times. The complementary pairing allowed the luminol to be immobilized on the surface of hemin@HKUST-1. The generation and consumption of short-lived reactive oxygen species were concentrated on the surface of the MOFs, which improves the chemiluminescence efficiency. The introduction of hemin@HKUST-1 and DNA solved the defects of chemiluminescence analysis. The chemiluminescence assay was able to detect lysozyme with linear range of 1.05 × 10−6 U∙mg−1 (6.00 × 10−13 mol∙L−1)–1.25 × 10−2 U∙mg−1 (7.14 × 10−9 mol∙L−1); the detection limit was 3.50 × 10−7 U∙mg−1 (2.00 × 10−13 mol∙L−1) (R2 = 0.99). The recovery of lysozyme in spiked saliva samples was 97.4–102.8%.
Keywords Carbon nanotube . Target recycling . Signal amplification . Protein . Enzyme . Catalytic effect . MOFs . Biosensor . DNA . Signal tag
Introduction
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-020-04254-2) contains supplementary material, which is available to authorized users. * Chuannan Luo [email protected] * Xueying Wang [email protected] 1
Key Laboratory of Interfacial Reaction & Sensing Analysis in Universities of Shandong, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022, People’s Republic of China
Magnetic nanomaterials are favored because of their characteristics of both nanomaterials and magnetic materials. The magnetic nanomaterials have been widely used in the fields of separation [1], adsorption [2, 3], drug targeted delivery [4, 5], and water treatment [6]. The chemical experiment process should follow the “5R” pr
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