Female BALB/c mice 6 to 8 weeks old were immunized with 30 to 50 μg of RBD recombinant protein mixed with TiterMax (Sigma-Aldrich, Canada) 3 times as we previously described [17 (link),58 (link),89 (link)]. The immunized spleen cells were then fused with mouse myeloma cells (P3X63Ag8.653, American Type Culture Collection), and hybridomas were selected by HAT medium (Sigma-Aldrich, Canada). Positive hybridomas were identified using enzyme-linked immunosorbent assay and subcloned through limit dilution. Hybridomas that secret antibodies were cultured in sera-free medium for large-scale antibody production. The mAbs were isolated/purified using Protein-G agarose beads (Thermo Fisher Scientific).
Ma X., Liang J., Zhu G., Bhoria P., Shoara A.A., MacKeigan D.T., Khoury C.J., Slavkovic S., Lin L., Karakas D., Chen Z., Prifti V., Liu Z., Shen C., Li Y., Zhang C., Dou J., Rousseau Z., Zhang J., Ni T., Lei X., Chen P., Wu X., Shaykhalishahi H., Mubareka S., Connelly K.A., Zhang H., Rotstein O, & Ni H. (2023). SARS-CoV-2 RBD and Its Variants Can Induce Platelet Activation and Clearance: Implications for Antibody Therapy and Vaccinations against COVID-19. Research, 6, 0124.
AgaroseAntibodies Antibody production Balb c miceCells Cultured medium Dilution Enzyme linked immunosorbent assay FemaleHybridomas Mabs Mouse Myeloma Protein g Recombinant protein SeraSpleenTitermax
Corresponding Organization :
Other organizations :
University of Toronto, St. Michael's Hospital, Canadian Blood Services, First Affiliated Hospital of Guangzhou Medical University, State Key Laboratory of Respiratory Disease, Guangzhou Medical University, Guangzhou University of Chinese Medicine, Sunnybrook Health Science Centre
RBD recombinant protein concentration (30 to 50 μg)
TiterMax adjuvant
dependent variables
Antibody production by hybridomas
control variables
Female BALB/c mice (6 to 8 weeks old)
Mouse myeloma cells (P3X63Ag8.653)
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
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