Vertical Flow Cellulose-Based Assays for SARS-CoV-2 Antibody Detection in Human Serum

We developed a SARS-CoV-2 antibody test using cellulose, an alternative membrane material, and a double-antigen sandwich format

Seunghyeon Kim; Yining Hao; Eric A. Miller; Dousabel M. Y. Tay; Emma Yee; Patthara Kongsuphol; Huan Jia; Megan McBee; Peter R. Preiser; Hadley D. Sikes

2021

Scholarcy highlights

  • Rapid and inexpensive serological tests for severe acute respiratory syndrome coronavirus-2 antibodies are essential to conduct large-scale seroprevalence surveys and can potentially complement nucleic acid or antigen tests at the point of care
  • Using nucleocapsid protein fused to cellulose-binding domain) as the capture reagent and biotinylated nucleocapsid protein with streptavidin horseradish peroxidase conjugate as the reporter reagent, we successfully demonstrated that target antibodies simultaneously associated with both NP-CBD and NP-biotin-SA-HRP were immobilized on the cellulosic test zone
  • The capture and reporter reagents used in this assay are NP fused to cellulose-binding domain and biotinylated NP, respectively
  • When a patient sample is mixed with the reagents, the reporting enzyme will be associated with CBD to form enzyme-antibodyCBD sandwich complexes as patient antibodies link the capture and reporter reagents
  • The sample-reagent mixture is applied to cellulose test zone, where the enzyme-antibody-CBD complexes are immobilized through specific binding between CBD and cellulose
  • We demonstrated that vertical-flow, cellulose-based tests can successfully detect SARS-CoV-2 antibodies in human serum using functionalized SARS-CoV-2 nucleocapsid protein as both capture and reporter reagents
  • We hope that this simple, rapid, and easy-to-scale approach to serological testing can be

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