신경괴사증바이러스(nervous necrosis virus, RGNNV genotype)에 대한단클론 항체 생산
Production of Monoclonal Antibodies Against Nervous Necrosis Virus (NNV, RGNNV genotype)
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초록
We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against nervous necrosis virus (NNV, RGNNV genotype). We established six hybridoma clones secreting MAbs against NNV antigen: 2B1, 2B11, 2C12, 13C1-1, 13C1-2 and 14D11. All six MAbs belonged to the IgG2a isotype with a kappa light chain and their reactivity recognized against the 41 kDa coat protein of NNV by Western blot analysis. The affinity constants of the six MAbs were measured by enzyme-linked immunosorbent assay (ELISA). All six MAbs reacted with two NNV isolates (SgNag05 and Gemunodo06), while no reactivity was observed with five know fish viruses, namely marine birnavirus, infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, hirame rhabdovirus, and infectious hematopoietic necrosis virus. Moreover, high ELISA optical density (OD) values (0.87-1.42) were observed in the brain tissues of NNV-infected sevenband grouper, while low OD values (less than 0.12) were recorded in the brain tissues of uninfected fish. These results suggest that these six MAbs are highly competent and useful for the detection of NNV with the RGNNV genotype.
Abstract
We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against nervous necrosis virus (NNV, RGNNV genotype). We established six hybridoma clones secreting MAbs against NNV antigen: 2B1, 2B11, 2C12, 13C1-1, 13C1-2 and 14D11. All six MAbs belonged to the IgG2a isotype with a kappa light chain and their reactivity recognized against the 41 kDa coat protein of NNV by Western blot analysis. The affinity constants of the six MAbs were measured by enzyme-linked immunosorbent assay (ELISA). All six MAbs reacted with two NNV isolates (SgNag05 and Gemunodo06), while no reactivity was observed with five know fish viruses, namely marine birnavirus, infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, hirame rhabdovirus, and infectious hematopoietic necrosis virus. Moreover, high ELISA optical density (OD) values (0.87-1.42) were observed in the brain tissues of NNV-infected sevenband grouper, while low OD values (less than 0.12) were recorded in the brain tissues of uninfected fish. These results suggest that these six MAbs are highly competent and useful for the detection of NNV with the RGNNV genotype.
참고문헌 (17)
- Cha SJ, "Phylogenetic analysis of betanodaviruses isolated from cultured fish in Korea", Dis Aquat Org, 77, pp.181~189 (2007)[DOI]
- Egusa S, "Infectious and parasitic diseases of fish and shellfish", pp.84~89 (2006), Life Science Publishing Co.
- Gomez DK, "Molecular detection of betanodavirus in wild marine fish populations in South Korea", J Vet Diagn Invest, 20, pp.38~44 (2008)[DOI]
- Gye HJ, "Purification of nervous necrosis virus (NNV) particles by anion-exchange chromatography", J Virol Method, 238, pp.21~28 (2016)[DOI]
- Gye HJ, "Heat-denaturation of conformational structures on nervous necrosis virus for generating neutralization antibodies", Aquaculture, 484, pp.65~70 (2018)[DOI]
- Jeong HN, "Production of monoclonal antibodies against viral hemorrhagic septicemia virus (VHSV, genotype IVa) from olive flounder", J Fish Pathol, 30, pp.149~154 (2017)[DOI]
- Johansen R, "Characterization of nodavirus and viral encephalophathy and retinopathy in farmed turbot, Scophthalmus maximus (L.)", J Fish Dis, 27, pp.591~601 (2004)[DOI]
- 김춘섭, "능성어 양식장에서의 viral nervous necrosis (VNN) 발생양상", 한국어병학회지, 25(2), pp.111~116 (2012), 한국어병학회[DOI]
- Kim YC, "Utilization of two serially connected PCR assay for detection and type-discrimination of betanodaviruses in shellfish", Korean federation of fisheries science and technology societies international conference 2017. Abstract, Ann Meet. J Fish Pathol, pp.195~ (2017)
- Laemmli UK, "Cleavage of structural proteins during the assembly of the head of bacteriophage T4", Nature, 227, pp.680~685 (1970)[DOI]
- Nam UH, "Monitoring of viruses in cultured walleye pollock Gadus chalcogrammmus", J Fish Pathol, 30, pp.1~9 (2017)[DOI]
- Nishizawa T, "Genomic classification of fish nodaviruses by molecular phylogenetic analysis of the coat protein genes", Appl Environ Microbiol, 63, pp.1633~1636 (1997)
- Oh MJ, "Field tests of poly(I:C) immunization with nervous necrosis virus (NNV) in sevenband grouper, Epinephelus septemfasciatus, (Thunberg)", J Fish Dis, 35, pp.187~191 (2012)[DOI]
- "Manual of diagnostic tests for aquatic animals. Viral encephalopathy and retinopathy" (2017)[URL]
- Schneemann A, "Virus taxonomy: Eighth report of the international committee on taxonomy of viruses", pp.2005, Elsevier
- Towbin H, "Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets; procedure and some applications", Proc Natl Acad Sci USA, 76, pp.4350~4354 (1979)[DOI]
- 원경미, "겨울철 양식 능성어의 바이러스성뇌망막증 (VER) 감염사례", 한국어류학회지, 29(3), pp.157~164 (2017), 한국어류학회