Development of a Single-Chain Variable Fragment of CR3022 for a Plasmonic-Based Biosensor Targeting the SARS-CoV-2 Spike Protein.

Two years after SARS-CoV-2 caused the first case of COVID-19, we are now in the "new normal" period, where people's activity has bounced back, followed by the easing of travel policy restrictions. The lesson learned is that the wide availability of accurate and rapid testing procedures is crucial to overcome possible outbreaks in the future. Therefore, many laboratories worldwide have been racing to develop a new point-of-care diagnostic test. To aid continuous innovation, we developed a plasmonic-based biosensor designed explicitly for portable Surface Plasmon Resonance (SPR). In this study, we designed a single chain variable fragment (scFv) from the CR3022 antibody with a particular linker that inserted a cysteine residue at the second position. It caused the linker to have a strong affinity to the gold surface through thiol-coupling and possibly become a ready-to-use bioreceptor toward a portable SPR gold chip without purification steps. The theoretical affinity of this scFv on spike protein was -64.7 kcal/mol, computed using the Molecular Mechanics Generalized Born Surface Area (MM/GBSA) method from the 100 ns molecular dynamics trajectory. Furthermore, the scFv was produced in Escherichia coli BL21 (DE3) as a soluble protein. The binding activity toward Spike Receptor Binding Domain (RBD) SARS-CoV-2 was confirmed with a spot-test, and the experimental binding free energy of -10.82 kcal/mol was determined using portable SPR spectroscopy. We hope this study will be useful in designing specific and low-cost bioreceptors, particularly early in an outbreak when the information on antibody capture is still limited.

Development of lateral flow assay based on anti-IBDV IgY for the rapid detection of Gumboro disease in poultry.

The poultry industry faces serious problems against infectious diseases, including Gumboro, which is caused by contagious bursal disease virus (IBDV). IBDV infects the bursa of Fabricius (BF), a lymphoid organ for controlling the B-cell maturation. Thus, it can trigger the secondary infection's vulnerability, leading to the high mortality and morbidity of the chicken. Moreover, managing the Gumboro post-outbreaks also requires considerable time and costs. Besides vaccination programs, the early detection of IBDV is vital as an outbreak control strategy. The most popular diagnostic tool is a lateral flow immunoassay or a rapid test that meets ASSURED (Affordable, Sensitive, Specific, User-friendly, Rapid and Robust, Equipment-free, and Deliverable to end-users) criteria. In this study, the lateral flow immunoassay was successfully developed based on anti-IBDV IgY as the bio receptor. Anti-IBDV IgY was successfully isolated from Isa Brown's egg yolk. The detection system showed an acceptable affinity against the inactivated IBDV sample (1.5 × 103 TCID50). In addition, it did not react with avian influenza and Newcastle disease viruses, demonstrating a good specificity of the test.