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10.1038/s41598-020-73960-x

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suck abstract from ncbi


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pmid33024203      Sci+Rep 2020 ; 10 (1): 16615
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  • Development of an antibody-dependent cellular cytotoxicity reporter assay for measuring anti-Middle East Respiratory Syndrome antibody bioactivity #MMPMID33024203
  • Cao J; Wang L; Yu C; Wang K; Wang W; Yan J; Li Y; Yang Y; Wang X; Wang J
  • Sci Rep 2020[Oct]; 10 (1): 16615 PMID33024203show ga
  • Middle East Respiratory Syndrome coronavirus (MERS-CoV) is a highly virulent pathogen that causes Middle East Respiratory Syndrome (MERS). Anti-MERS-CoV antibodies play an integral role in the prevention and treatment against MERS-CoV infections. Bioactivity is a key quality attribute of therapeutic antibodies, and high accuracy and precision are required. The major methods for evaluating the antiviral effect of antiviral antibodies include neutralization assays using live viruses or pseudoviruses are highly variable. Recent studies have demonstrated that the antibody-dependent cellular cytotoxicity (ADCC) activity of antiviral antibodies is more consistent with the virus clearance effect in vivo than neutralization activity. However, no reports evaluating the ADCC activity of anti-MERS antibodies have been published to date. Here, we describe the development of a robust and reliable cell-based reporter gene assay for the determination of ADCC activity of anti-MERS antibodies using 293T/MERS cells stably expressing the spike protein of MERS-CoV (MERS-S) as target cells and the engineered Jurkat/NFAT-luc/FcgammaRIIIa stably expressing FcgammaRIIIA and NFAT reporter gene as effector cells. According to the ICH-Q2 analytical method guidelines, we carefully optimized the experimental conditions and assessed the performance of our assay. In addition, we found that the ADCC activity of afucosylated anti-MERS antibodies is higher than their fucosylated counterparts. The establishment of this ADCC determination system provides a novel method for evaluating the bioactivity of anti-MERS antibodies and improving ADCC activity through modification of N-glycosylation of the Fc segment.
  • |Antibodies, Neutralizing/immunology[MESH]
  • |Antibodies, Viral/*analysis/immunology[MESH]
  • |Antibody-Dependent Cell Cytotoxicity/*immunology[MESH]
  • |Coronavirus Infections/*immunology/virology[MESH]
  • |Cytotoxicity Tests, Immunologic/*methods[MESH]
  • |Genes, Reporter[MESH]
  • |HEK293 Cells[MESH]
  • |Humans[MESH]
  • |Jurkat Cells[MESH]
  • |Luciferases/genetics[MESH]
  • |Middle East Respiratory Syndrome Coronavirus/genetics/*immunology/*metabolism[MESH]
  • |NFATC Transcription Factors/genetics[MESH]
  • |Receptors, IgG/genetics/immunology[MESH]
  • |Response Elements[MESH]
  • |Spike Glycoprotein, Coronavirus/metabolism[MESH]


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