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http://dx.doi.org/10.5483/BMBRep.2019.52.6.185

Generation and characterization of a monoclonal antibody against MERS-CoV targeting the spike protein using a synthetic peptide epitope-CpG-DNA-liposome complex  

Park, Byoung Kwon (Center for Medical Science Research, College of Medicine, Hallym University)
Maharjan, Sony (Center for Medical Science Research, College of Medicine, Hallym University)
Lee, Su In (Center for Medical Science Research, College of Medicine, Hallym University)
Kim, Jinsoo (Department of Microbiology, College of Medicine, Hallym University)
Bae, Joon-Yong (Department of Microbiology, College of Medicine, and the Institute for Viral Diseases, Korea University)
Park, Man-Seong (Department of Microbiology, College of Medicine, and the Institute for Viral Diseases, Korea University)
Kwon, Hyung-Joo (Center for Medical Science Research, College of Medicine, Hallym University)
Publication Information
BMB Reports / v.52, no.6, 2019 , pp. 397-402 More about this Journal
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) uses the spike (S) glycoprotein to recognize and enter target cells. In this study, we selected two epitope peptide sequences within the receptor binding domain (RBD) of the MERS-CoV S protein. We used a complex consisting of the epitope peptide of the MERS-CoV S protein and CpG-DNA encapsulated in liposome complex to immunize mice, and produced the monoclonal antibodies 506-2G10G5 and 492-1G10E4E2. The western blotting data showed that both monoclonal antibodies detected the S protein and immunoprecipitated the native form of the S protein. Indirect immunofluorescence and confocal analysis suggested strong reactivity of the antibodies towards the S protein of MERS-CoV virus infected Vero cells. Furthermore, the 506-2G10G5 monoclonal antibody significantly reduced plaque formation in MERS-CoV infected Vero cells compared to normal mouse IgG and 492-1G10E4E2. Thus, we successfully produced a monoclonal antibody directed against the RBD domain of the S protein which could be used in the development of diagnostics and therapeutic applications in the future.
Keywords
B cell epitope; Lipoplex (O); MERS-CoV; Monoclonal antibody; Spike protein;
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Times Cited By KSCI : 4  (Citation Analysis)
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1 Park BK, Lee SI, Bae JY et al (2018) Production of a Monoclonal Antibody targeting the M protein of MERSCoV for detection of MERS-CoV using a synthetic peptide epitope formulated with a CpG-DNA-liposome complex. Int J Pept Res Ther https://doi.org/10.1007/s10989-018-9731-8
2 Salazar G, Zhang N, Fu TM and An Z (2017) Antibody therapies for the prevention and treatment of viral infections. NPJ Vaccines 2, 19   DOI
3 Park BK, Park JY, Kim TH et al (2018) Production of an anti-TM4SF5 monoclonal antibody and its application in the detection of TM4SF5 as a possible marker of a poor prognosis in colorectal cancer. Int J Oncol 53, 275-285
4 Wu G, Maharjan S, Kim D et al (2018) A Novel monoclonal antibody targets mucin1 and attenuates growth in pancreatic cancer model. Int J Mol Sci 19, 2004   DOI
5 Park BK, Gautam A, Maharjan S et al (2017) Production of anti-c-Myc monoclonal antibody inhibiting DNA binding of c-Myc and max dimer by epitope peptide-CpGDNA-liposome complex without carriers. Int J Pept Res Ther https://doi.org/10.1007/s10989-017-9649-6
6 Wu G, Kim D, Kim JN et al (2018) A Mucin1 C-terminal subunit-directed monoclonal antibody targets overexpressed Mucin1 in breast cancer. Theranostics 8, 78-91   DOI
7 Kim D, Kwon S, Rhee JW et al (2011) Production of antibodies with peptide-CpG-DNA-liposome complex without carriers. BMC Immunol 12, 29   DOI
8 Maharjan S, Park BK, Lee SI et al (2018) Gomisin G inhibits the growth of triple-negative breast cancer cells by suppressing AKT phosphorylation and decreasing cyclin D1. Biomol Ther (Seoul) 26, 322-327   DOI
9 Maharjan S, Park BK, Lee SI et al (2018) Gomisin G suppresses the growth of colon cancer cells by attenuation of AKT phosphorylation and arrest of cell cycle progression. Biomol Ther (Seoul) https://doi.org/10.4062/biomolther.2018.054
10 Jo HS, Eum WS, Park EY et al (2017) Effects of PEP-1-FK506BP on cyst formation in polycystic kidney disease. BMB Rep 50, 460-465   DOI
11 Lee EW, Oh W, Song HP and Kim WK (2017) Phosphorylation of p53 at threonine 155 is required for Jab1-mediated nuclear export of p53. BMB Rep 50, 373-378   DOI
12 Yokoyama WM, Christensen M, Santos GD and Miller D (2006) Production of monoclonal antibodies. Curr Protoc Immunol 102, Unit 2.5
13 Durai P, Batool M, Shah M and Choi S (2015) Middle East respiratory syndrome coronavirus: transmission, virology and therapeutic targeting to aid in outbreak control. Exp Mol Med 47, e181   DOI
14 Zumla A, Chan JF, Azhar EI et al (2016) Coronaviruses - drug discovery and therapeutic options. Nat Rev Drug Discov 15, 327-347   DOI
15 Du L, Yang Y, Zhou Y et al (2017) MERS-CoV spike protein: a key target for antivirals. Expert Opin Ther Targets 21, 131-143   DOI
16 Mackay IM and Arden KE (2015) MERS coronavirus: diagnostics, epidemiology and transmission. Virol J 12, 222   DOI
17 Zaki AM, van Boheemen S, Bestebroer TM et al (2012) Isolation of a novel coronavirus from a man with pneumonia in Saudi Arabia. N Engl J Med 367, 1814-1820   DOI
18 Omrani AS, Al-Tawfiq JA and Memish ZA (2015) Middle East respiratory syndrome coronavirus (MERS-CoV): animal to human interaction. Pathog Glob Health 109, 354-362   DOI
19 World Health Organization, Middle East respiratory syndrome coronavirus (MERS-CoV), 2018. (http://www.who.int/emergencies/mers-cov/en/)
20 Ki M (2015) 2015 MERS outbreak in Korea: hospital-tohospital transmission. Epidemiol Health 37, e2015033   DOI
21 Lambour J, Naranjo-Gomez M, Piechaczyk M and Pelegrin M (2016) Converting monoclonal antibody-based immunotherapies from passive to active: bringing immune complexes into play. Emerg Microbes Infect 5, e92   DOI
22 Scott AM, Allison JP and Wolchok JD (2012) Monoclonal antibodies in cancer therapy. Cancer Immun 12, 14
23 Wang Q, Wong G, Lu G et al (2016) MERS-CoV spike protein: Targets for vaccines and therapeutics. Antiviral Res 133, 165-177   DOI
24 Kim D, Kwon HJ and Lee Y (2011) Activation of Toll-like receptor 9 and production of epitope specific antibody by liposome-encapsulated CpG-DNA. BMB Rep 44, 607-612   DOI
25 Gao J, Lu G, Qi J et al (2013) Structure of the fusion core and inhibition of fusion by a heptad repeat peptide derived from the S protein of Middle East respiratory syndrome coronavirus. J Virol 87, 13134-13140   DOI
26 Wang N, Shi X, Jiang L et al (2013) Structure of MERS-CoV spike receptor-binding domain complexed with human receptor DPP4. Cell Res 23, 986-993   DOI
27 Raj VS, Mou H, Smits SL et al (2013) Dipeptidyl peptidase 4 is a functional receptor for the emerging human coronavirus-EMC. Nature 495, 251-254   DOI
28 Park BK, Choi SH, Kim YE et al (2015) Monoclonal antibodies against the human respiratory syncytial virus obtained by immunization with epitope peptides and CpG-DNA-liposome complex. Monoclon Antib Immunodiagn Immunother 34, 101-109   DOI
29 Rhee JW, Kim D, Park et al (2012) Immunization with a hemagglutinin-derived synthetic peptide formulated with a CpG-DNA-liposome complex induced protection against lethal influenza virus infection in mice. PLoS One 7, e48750   DOI