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A functionalized Sup35NM nanofibril-assisted oriented antibody capture in lateral flow immunoassay for sensitive detection of dengue type II NS1

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Abstract

Rapid and sensitive dengue non-structural protein 1 (NS1) detection assay is essential for the treatment of disease and currently releases high medical cost burdens. To address the limitations of conventional LFIA strips, we have developed an improved Sup35NM-Z-based LFIA that immobilizes antibodies on cellulose membranes in an orientated manner to increase the sensitivity of LFIA strips. A dual-functional Sup35NM nanofibril was fabricated by fusion with the antibody binding domain; resultant nanofibril from the amyloid Sup35NM was sprayed on the T-line to orientate the capture antibody and produces fluorescence signals. Antibody binding analysis showed that self-assembly of the Sup35NM monomer does not affect the binding activity of the Z-domain with the antibody. The NS1 for DENV-2 infection was chosen as a model target antigen to assess the feasibility of the Sup35NM-Z-domain-based LFIA platform. Under optimal conditions, the Sup35NM-Z-domain-based LFIA detected NS1 within 15 min with a detection limit of 1.29 ng/ml, while the detection limit of traditional LFIA with the same concentration of anti-NS1-Ab1 on the T-line by conventional physical adsorption was 2.20 ng/ml, 1.7 times higher than that of Sup35NM-Z-domain-based LFIA. As compared to traditional LFIAs, the Sup35NM-Z-based LFIA had a wide detection range of 1.29–625 ng/mL. The LFIA's clinical performance in identifying NS1 was also assessed using 15 clinical samples. The LFIA accurately recognized positive and negative samples, equal to 86.7% accuracy. The developed Sup35NM-Z-domain-based LFIA in this study offers great potential for the identification of target markers because of its greatly improved sensitivity and wider detection range.

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Funding

This work was financially supported by Science and Technology Program of Guangzhou, China (202205110007).

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Authors and Affiliations

  1. School of Biology and Biological Engineering, South China University of Technology, Guangzhou, 510006, China

    Ting Qin, Yi Ma & Jufang Wang

  2. National Engineering Laboratory of Rapid Diagnostic Tests, Guangzhou Wondfo Biotech Co., Ltd., Guangzhou, 510663, China

    Peidian Wu, Qiankun Zhang & Keren Kang

  3. Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, South China University of Technology, Guangzhou, 510006, China

    Yi Ma & Jufang Wang

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  1. Ting Qin
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  2. Peidian Wu
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  3. Qiankun Zhang
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Contributions

TQ: investigation, methodology, data curation, experimentation, writing—original draft. PW: investigation, resources. QZ and KK: formal analysis, resources. YM: formal analysis, investigation, supervision, project administration. JW: conceptualization; writing, review and editing; supervision; project administration; funding acquisition. All authors read and approved the final manuscript.

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Correspondence to Jufang Wang.

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Highlights

The Sup35NM-Z-domain can self-assemble into nanofibers driven by Sup35NM.

The Sup35NM-Z-domain immobilizes antibodies on cellulose membranes in an orientated manner.

The Sup35NM-Z-based LFIA configuration does not need expensive secondary antibodies and reduces the chemical labeling step of the antibody.

The proposed method for NS1 detection achieves improved sensitivity with a LOD of 1.29 ng/ml.

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Qin, T., Wu, P., Zhang, Q. et al. A functionalized Sup35NM nanofibril-assisted oriented antibody capture in lateral flow immunoassay for sensitive detection of dengue type II NS1. Microchim Acta 191, 39 (2024). https://doi.org/10.1007/s00604-023-06109-y

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