51. Immunoglobulin fragment F(ab’)2 against RBD potently neutralizes SARS-CoV-2 in vitro
- Author
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Jing Zhang, Fang Lijuan, Xiaoyan Pan, Yuan Sun, Rui Gong, Ze Chen, Gengfu Xiao, Tiejiong Fan, Pengfei Zhou, Weijuan Shang, Shaojuan Zhao, Jian Shi, Xiaoyue Shi, Xiaming Jiang, and Yan Wu
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0301 basic medicine ,viruses ,Antibodies, Viral ,law.invention ,law ,Chlorocebus aethiops ,Receptors, Immunologic ,Receptor ,Neutralizing antibody ,skin and connective tissue diseases ,Mice, Inbred BALB C ,biology ,Chemistry ,Chinese hamster ovary cell ,neutralizing antibody ,virus diseases ,immunoglobulin fragment ,Spike Glycoprotein, Coronavirus ,Recombinant DNA ,Female ,Antibody ,receptor-binding domain ,Coronavirus Infections ,Protein Binding ,030106 microbiology ,Pneumonia, Viral ,Article ,03 medical and health sciences ,Betacoronavirus ,Immune system ,In vivo ,Neutralization Tests ,Virology ,Animals ,Humans ,Pandemics ,Vero Cells ,EC50 ,Pharmacology ,Antiserum ,SARS-CoV-2 ,fungi ,COVID-19 ,Fragment crystallizable region ,Antibodies, Neutralizing ,In vitro ,body regions ,030104 developmental biology ,biology.protein ,HeLa Cells - Abstract
COVID-19, which is caused by the emerging human coronavirus SARS-CoV-2, has become a global pandemic that poses a serious threat to human health. To date, no vaccines or specific antiviral drugs have been approved for the treatment of this disease in clinic. Herein, therapeutic antibodies for SARS-CoV-2 were obtained from hyperimmune equine plasma. First, a recombinant SARS-CoV-2 spike protein receptor-binding domain (RBD) was obtained in gram-level quantities through high-cell density fermentation of Chinese hamster ovary cells. Then, the binding of the RBD to the SARS-CoV-2 receptor, human angiotensin-converting enzyme 2, was verified by several biochemical methods. The efficacy of the RBD in triggering antibody response in vivo was subsequently tested in both mice and equines, and the results showed that the RBD triggered high-titer neutralizing antibody production in vivo. Immunoglobulin F(ab’)2 fragments were prepared from equine antisera via removal of the Fc region from the immunoglobulins. Finally, a neutralization test with live virus demonstrated that RBD-specific F(ab’)2 inhibited SARS-CoV-2 with an EC50 of 0.07 μg/ml and an EC80 of 0.18 μg/ml, showing a potent inhibitory effect on SARS-CoV-2. These results highlight RBD-specific equine immunoglobulin F(ab’)2 fragment as a candidate for the treatment of SARS-CoV-2., Highlights • SARS-CoV-2 RBD triggered strong antibody responses both in mice and equines. • High neutralizing-titer antisera were obtained by immunizing equine with CHO cell-expressed RBD protein. • RBD-specific immunoglobulin F(ab’)2 fragments potently neutralized SARS-CoV-2 in vitro.
- Published
- 2020
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