Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

A Paper-based Immunofluorescent Device for the Detection of SARS-CoV-2 Humanized Antibody

Version 1 : Received: 7 July 2021 / Approved: 7 July 2021 / Online: 7 July 2021 (13:18:33 CEST)

How to cite: Kasetsirikul, S.; Umer, M.; Soda, N.; Sreejith, K.R.; Shiddiky, M.J.A.; Nguyen, N. A Paper-based Immunofluorescent Device for the Detection of SARS-CoV-2 Humanized Antibody. Preprints 2021, 2021070181 (doi: 10.20944/preprints202107.0181.v1). Kasetsirikul, S.; Umer, M.; Soda, N.; Sreejith, K.R.; Shiddiky, M.J.A.; Nguyen, N. A Paper-based Immunofluorescent Device for the Detection of SARS-CoV-2 Humanized Antibody. Preprints 2021, 2021070181 (doi: 10.20944/preprints202107.0181.v1).

Abstract

We report on an immunofluorescent paper-based assay for the detection of severe acute respiratory symptom coronavirus 2 (SARS-CoV-2) humanized antibody. The paper-based device was fabricated by using lamination technique for easy and optimized handling. Our approach utilises a two-step strategy that involves (i) initial coating of the paper-electrode with recombinant SARS-CoV-2 nucleocapsid antigen to capture the target SARS-CoV-2 specific antibodies, and (ii) subsequent detection of SARS-CoV-2 antibodies using fluorophore-conjugated IgG antibody. The fluorescence readout was observed with fluorescence microscopy. The images were processed and quantified using a MATLAB program. The assay can selectively detect SARS-CoV-2 humanized antibodies spiked in PBS and healthy human serum samples with the relative standard deviation of approximately 6.4% (for n = 3). It has broad dynamic ranges (1 ng to 50 ng/µL in PBS and 5 to 100 ng/µL in human serum samples) for SARS-CoV-2 humanized antibodies with the detection limits of 2 ng/µL (0.025 IU/mL) and 10 ng/µL (0.125 IU/mL) in PBS and human serum samples, respectively. We believe that our assay has the potential to be used as a simple, rapid, and inexpensive paper-based diagnostic device with a portable fluorescent reader to provide point-of-care diagnosis. This assay can be used for rapid examination of a large batch of samples toward clinical screening of SARS-CoV-2 specific antibodies as a confirmed infected active case or to evaluate the immune response to a SARS-CoV-2 vaccine.

Subject Areas

SARS-CoV-2 detection; Immunofluorescence; Paper-based diagnostic device

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