Removal of 3C protease from the 3ABC improves expression, solubility, and purification of the recombinant 3AB of foot-and-mouth disease virus

Salem, Reda; El-Kholy, Alaa A.; Waly, Fatma R.; Khaled, Radwa; Elmenofy, Wael

doi:10.1007/s11262-020-01815-8

Cited by 2 publications

(1 citation statement)

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“…Infected cells were cultured in T-flasks for small-scale production, and culture supernatants, as well as infected cells, were harvested daily at 3–7 days post-infection (DPI). Each harvest was subjected to sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analyses (El-Gaied et al 2017 ; Salem et al 2021 ), to verify integrity and immunoreactivity of the expressed proteins, as well as their peak-expression times.…”

Section: Methodsmentioning

confidence: 99%

A gold nanoparticles-based lateral flow assay utilizing baculovirus expressed recombinant nucleocapsid and receptor binding domain proteins for serodetection of IgG and IgM against SARS-CoV-2

et al. 2022

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Serological assays for SARS-CoV-2 are being utilized at an exponential rate for surveillance programs. This enterprise was designed to develop and validate a qualitative immunochromatographic test, via the Lateral Flow Assay (LFA), for detection of immunoglobulins M and G (IgM and IgG) against both nucleocapsid (N) and the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2. Both targeted proteins were cloned and expressed in baculovirus expression system utilizing insect cells Sf9. The recombinant RBD and N proteins were purified and conjugated with gold nanoparticles (AuNPs) to set up the coating antigens pad. Both anti-human IgG and IgM were dispensed on nitrocellulose membrane to capture human antibodies in serum samples. A home-made dispensing system was developed to draw identical test and control lines. The validity of the developed LFA was verified by testing serum samples from 103 convalescent COVID-19 patients who were PCR positive for SARS-CoV-2 along with 28 control serum samples. The developed strips showed distinctive bands for IgM and IgG of both proteins (RBD and N) in positive samples. The sensitivity of RBD-based LFA was 70.9% and 39.8% for IgG and IgM, respectively, with a specificity of 100% for both. The N-based LFA exhibited a sensitivity of 73.8% and 35.9% for IgG and IgM, respectively, while its specificity was 75% and 100% for IgG and IgM, respectively. Our developed LFA could afford a tool for surveillance programs in low-resource countries. Moreover, it might be functional for rapid and inexpensive monitoring of the anti-SARS-CoV-2 antibodies in the sera of vaccinated individuals.

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Section: Methodsmentioning

confidence: 99%