Baculovirus expression system

Manufactured by Sino Biological

Sourced in United States

The baculovirus expression system is a widely used method for the production of recombinant proteins. It utilizes insect cells and baculoviruses as the expression system. The core function of this system is to enable the efficient expression and purification of a variety of recombinant proteins.

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Lab products found in correlation

Hrp conjugated goat anti human igg, by Southern Biotech (2 mentions) O phenylenediamine dihydrochloride peroxidase substrate, by Merck Group (1 mentions) O phenylenediamine, by Merck Group (1 mentions)

3 protocols using baculovirus expression system

Recombinant SARS-CoV-2 and Influenza Nucleoproteins

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Recombinant nucleoprotein
(N) of SARS-CoV-2 (2019-nCov), Influenza A (A/Wisconsin/588/2019–A/Victoria/2570/2019),
and Influenza B (B/Phuket/3073/2013) was produced in insect cells
with a baculovirus expression system (Sino Biological, Beijing, China).
The SARS-CoV-2 sample preparation protocol was automated using an
Andrew Alliance Pipette+ and Shaker+ connected device (Waters Corporation,
Milford, MA, USA) both operated via the OneLab platform.

Van Puyvelde B., Van Uytfanghe K., Van Oudenhove L., Gabriels R., Van Royen T., Matthys A., Razavi M., Yip R., Pearson T., Drouin N., Claereboudt J., Foley D., Wardle R., Wyndham K., Hankemeier T., Jones D., Saelens X., Martens G., Stove C.P., Deforce D., Martens L., Vissers J.P., Anderson N.L, & Dhaenens M. (2022). Cov2MS: An Automated and Quantitative Matrix-Independent Assay for Mass Spectrometric Measurement of SARS-CoV-2 Nucleocapsid Protein. Analytical Chemistry, 94(50), 17379-17387.

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Anti-Vimentin Antibody ELISA Assay

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Anti-vimentin antibodies were measured by a standard ELISA against recombinant human vimentin purified from a baculovirus expression system (SinoBiological, Wayne, PA, USA). Vimentin in bicarbonate buffer (5 μg/mL), pH 9.4, was coated on ELISA plates overnight at 4 °C. All sera were tested at a 1:100 dilution. Serial dilutions of a pooled serum sample were used as a calibrator for each plate. Bound antibody was detected with HRP-conjugated goat anti-human IgG (SouthernBiotech, Birmingham AL, USA), and the enzyme activity was determined with an o-phenylenediamine dihydrochloride (OPD) peroxidase substrate (Sigma-Aldrich, St. Louis, MO, USA). The reaction was stopped with 2.5 N sulfuric acid. Results are presented as absorbance at 490 nm. The mean O.D. +2SD of the control group was used as a cut-off to determine the prevalence of anti-vimentin antibodies.

Bagavant H., Cizio K., Araszkiewicz A.M., Papinska J.A., Garman L., Li C., Pezant N., Drake W.P., Montgomery C.G, & Deshmukh U.S. (2022). Systemic immune response to vimentin and granuloma formation in a model of pulmonary sarcoidosis. Journal of Translational Autoimmunity, 5, 100153.

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ELISA-Based Vimentin Antibody Quantification

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An ELISA-based assay was used to determine antibodies to purified recombinant human vimentin generated in a baculovirus expression system (Sino Biologicals, Wayne, PA, USA) as previously described [15 (link)]. Briefly, vimentin in bicarbonate buffer (5μg/ml) was coated on ELISA plates overnight at 4°C. All sera were tested at a 1:100 dilution. In each plate, serial dilutions of pooled serum samples positive for anti-vimentin were used as calibrators. Bound antibody was detected with HRP- conjugated goat anti-human IgG (Southern Biotech, Birmingham, AL, USA). O-phenylenediamine (Sigma Aldrich, St. Louis, MO, USA) was used as a substrate to determine enzyme activity, and the reaction was stopped with 2.5N sulfuric acid. The results are presented as absorbance at 490nm. The cut-off for anti-vimentin positivity was set at mean ± 2SD of the non-SjD sicca and healthy control sera to evaluate the association between the presence of anti-vimentin antibody and SjD.

Bagavant H., Araszkiewicz A.M., Rasmussen A., Pezant N., Montgomery C., Scofield R.H., Farris D., Lessard C.J, & Deshmukh U.S. (2023). Anti-Vimentin antibodies are associated with higher severity of Sjögren’s disease. Clinical immunology (Orlando, Fla.), 247, 109243.

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