Investigating Membrane Morphology and Quantity of Immobilized Protein for the Development of Lateral Flow Immunoassay

Ahmad, A.L.; Low, Siew Chun; Shukor, Syamsul Rizal Abd; Ismail, Asma

doi:10.1080/15321819.2011.591479

Cited by 6 publications

(2 citation statements)

References 21 publications

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“…First, each VFI prototype was constructed using 0.1 μm pore size NC membranes, as the signal intensity can be enhanced with this relatively small pore size. The reasoning for this is that a smaller pore NC membrane has a larger surface area for Ag–Ab binding than a larger pore. , Indeed, our previous studies also confirmed that the 0.1 μm pore size enhanced the signal intensity approximately twofold relative to a 5 μm pore size . Previously, we built a theoretical model to simulate the antigen–antibody binding process inside the nitrocellulose membrane to identify the suitable membrane pore size and sample flow rate to improve the detection sensitivity.…”

Section: Resultsmentioning

confidence: 65%

Critical Comparison between Large and Mini Vertical Flow Immunoassay Platforms for Yersinia Pestis Detection

Devadhasan

Chen

et al. 2021

Anal. Chem.

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Yersinia pestis is a Gram-negative bacterium that is the causative agent of plague and is widely recognized as a potential biological weapon. Due to the high fatality rate of plague when diagnosis is delayed, the development of rapid, sensitive, specific, and cost-effective methods is needed for its diagnosis. The Y. pestis low calcium response V (LcrV) protein has been identified as a potential microbial biomarker for the diagnosis of plague. In this paper, we present a highly sensitive, paper-based, vertical flow immunoassay (VFI) prototype for the detection of LcrV and the diagnosis of plague. An antigen-capture assay using monoclonal antibodies is employed to capture and detect the LcrV protein, using a colorimetric approach. In addition, the effect of miniaturizing the VFI device is explored based on two different sizes of VFI platforms, denoted as “large VFI” and “mini VFI.” Also, a comparative analysis is performed between the VFI platform and a lateral flow immunoassay (LFI) platform to exhibit the improved assay sensitivity suitable for point-of-care (POC) diagnostics. The analytical sensitivity or limit of detection (LOD) in the mini VFI is approximately 0.025 ng/mL, that is, 10 times better than that of the large VFI platform or 80 times over a standard lateral flow configuration. The low LOD of the LcrV VFI appears to be highly suitable for testing clinical samples and potentially diagnosing plague at earlier time points. In addition, optimization of the gold nanoparticle (AuNP) concentration, nanomaterial plasmonic properties, and flow velocity analysis could improve the performance of the VFI. Furthermore, we developed automated image analysis software that shows potential for integrating the diagnostic system into a smartphone. These methods and findings demonstrate that the VFI platform is a highly sensitive device for detecting the LcrV and potentially many other biomarkers.

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Section: Resultsmentioning

confidence: 65%

Critical Comparison between Large and Mini Vertical Flow Immunoassay Platforms for Yersinia Pestis Detection

Devadhasan

Chen

et al. 2021

Anal. Chem.

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“…The protein adsorptive value increases with the increase of polymer concentration. Thin film with higher polymer content leads to a better protein‐polymer binding affinity, due to the electrostatic interaction . Therefore, the highest protein binding was observed at 1359 ± 101.07 μg cm −3 , which was prepared by using 28 wt % of nylon polymer, whereas, only 994 ± 47.05 μg cm −3 protein binding was observed for nylon film with 16 wt % of polymer (N‐16 film).…”

Section: Resultsmentioning

confidence: 99%

Morphological characteristics of polymeric nylon‐6 film as biological recognition interface for electrochemical immunosensor application

Shaimi

Low

2018

J of Applied Polymer Sci

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The present study elucidates the morphology characteristic of nylon‐6 as protein immobilization film and their influences in biological recognition interface for a single‐use electrochemical immunosensor. A basic requirement of this polymer platform is to allow the testing analyte [conductive polyaniline‐Fe2O3‐glutaraldehyde‐conjugated‐N‐hydroxysuccinimidobiotin (PANI‐Fe2O3‐GA‐b‐NHS)] to access to the immobilized capture reagent [bovine serum albumin (BSA)] on a surface of the nylon‐6 film. As an immunoassay, the biochemical reaction between PANI‐Fe2O3‐GA‐b‐NHS and BSA takes place on a surface of the nylon‐6 support film, which then translated into measurable resistance signal through pulse‐mode measurement. The in‐house developed nylon‐6 film, N‐16B, with 16 wt % nylon polymer and methanol as nonsolvent had demonstrated the fastest lateral wicking speed (1.07 mm s−1) and excellent protein immobilization capacity (1650 ± 85.84 μg cm−3), leading to the higher electrical current signal with a minimal resistance of electrochemical responses (0.57 ± 0.04 MΩ). This work reflects the importance of the physical characteristics of the nylon‐6 film that determines the sensitivity and effectiveness of an immunosensor. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018, 135, 46741.

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Rapid mastitis detection assay on porous nitrocellulose membrane slides

2013

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We have developed a rapid mastitis detection test based on the immobilization of tag-specific antibody molecules, the binding of double-tagged amplicons, and as a secondary signal a conjugate of black carbon nanoparticles having molecules of a fusion protein of neutrAvidin and alkaline phosphatase at their surface. The antibodies were inkjet printed onto three different nitrocellulose membrane slides, Unisart (Sartorius), FAST (GE Whatman), and Oncyte-Avid (Grace-Biolabs), and the final assay signals on these slides were compared. The blackness of the spots was determined by flatbed scanning and assessment of the pixel gray volume using TotalLab image analysis software. The black spots could be easily read by the naked eye. We successfully demonstrated the detection of specific amplicons from mastitis-causing pathogens in less than 3 h. Using a similar protocol, we also showed that it was possible to detect specific amplicons from four different mastitis-causing pathogens (six strains) on the same pad. The influence of two different printing buffers, phosphate-buffered saline (pH 7.4) and carbonate buffer (pH 9.6), on the functionality of the primary antibodies was also compared.

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Investigating Membrane Morphology and Quantity of Immobilized Protein for the Development of Lateral Flow Immunoassay

Cited by 6 publications

References 21 publications

Critical Comparison between Large and Mini Vertical Flow Immunoassay Platforms for Yersinia Pestis Detection

Critical Comparison between Large and Mini Vertical Flow Immunoassay Platforms for Yersinia Pestis Detection

Morphological characteristics of polymeric nylon‐6 film as biological recognition interface for electrochemical immunosensor application

Rapid mastitis detection assay on porous nitrocellulose membrane slides

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