Description:Mouse monoclonal antibody to human hepatitis C virus (HCV) core protein

Purification:Protein G affinity purified

Product Type:Primary Antibody

Target Protein:Human hepatitis C virus (HCV) core protein

Immunogen:Synthetic peptides derived from HCV core protein

Fusion Myeloma:Sp2/0-Ag14

Specificity:React with recombinant core protein C + envelope protein M (residues 1-142 on HCV polyprotein) and synthetic core protein C (residues 1-61 on HCV polyprotein), and recombinant chimeric HCV polyprotein (60kDa).

Species Reactivity:Human hepatitis C virus, others not tested

Cross-Reactivity:No cross reaction with recombinant NS-3 protein (residues 1252-1477), synthetic NS-3 protein (residues 1378 -1458) and synthetic NS-4a protein (residues 1689 -1735). It recognizes different antigenic determinants of HCV core protein.

Host / Isotype:Mouse, IgG1 Kappa

Formulation:Lyophilized from a solution in 0.01M PBS, pH 7.0

Reconstitution:Double distilled water is recommended to adjust the final concentration to 1.00mg/mL

Storage: Store at -20^oC

Research Area:Virology

Background:

Hepatitis C virus (HCV) causes chronic hepatitis and liver cirrhosis in human through blood and body fluid transmission. HCV has a positive sense single RNA genome enclosed in the nucleocapsid made of Core Protein (Capsid Protein). The nucleocapsid is covered by an envelope made of lipoproteins (E1 and E2). The 9.6 kb HCV genome has a single open-reading frame, which is to be translated into a single polyprotein. HCV viral proteins are produced after processing the polyprotein. Genes for core protein and envelop proteins are located adjacently at the 5’-end of HCV genome, followed by genes for non-structural proteins including NS2, NS3, NS4A, NS4B, NS5, NS5A and NS5B.

Application:

ELISA:The mAb reacts with human hepatitis C virus core protein, and doesn’t react with HCV non-structure protein.

Western Blot:The mAb, when used at concentration of 0.1-0.5μg/mL, will allow visualization of 0.1 μg/lane of recombinant core protein C + envelope protein M, 0.5 μg /lane synthetic core protein C, and 0.1μg /lane recombinant 60kDa chimeric HCV polyprotein. The mAb works on blots transferred from both reducing and non-reducing PAGE gel. The mAb recognizes in-vitro translated HCV core protein.

Immunofluorescent analysis (see image):Confocal immunofluorescent analysis of human hepatitis C virus core protein using anti-HCV core protein mAb clone B2. (Photo provided by Dr. Rodney S. Russell, Faculty of Medicine, Memorial University of Newfoundland).

References:

1. Candice Jackel-Cram et al. (2010) Hepatitis C virus genotype-3a core protein enhances sterol regulatory element-binding protein-1 activity through the phosphoinositide 3-kinase–Akt-2 pathway. Journal of General Virology, 91:1388–1395.

2. Tiziana Vescovo et al. Autophagy Protects Cells From HCV-Induced Defects in Lipid Metabolism. Gastroenterology Volume 142, Issue 3, Pages 644–653.e3, March 2012.

3. Judith M. Gottwein et al. Robust Hepatitis C Genotype 3a Cell Culture Releasing Adapted Intergenotypic 3a/2a (S52/JFH1) Viruses. Gastroenterology Volume 133, Issue 5, Pages 1614–1626, November 2007.

4. Troels K.H. Scheel et al. Recombinant HCV Variants With NS5A From Genotypes 1–7 Have Different Sensitivities to an NS5A Inhibitor but Not Interferon-α. Gastroenterology Volume 140, Issue 3, Pages 1032–1042.e6, March 2011.

5. Takeya Tsutsumi et al. Proteomics analysis of mitochondrial proteins reveals overexpression of a mitochondrial protein chaperon, prohibitin, in cells expressing hepatitis C virus core protein. Hepatology Volume 50, Issue 2, pages 378–386, August 2009.

6. Tsutsumi T et al. Interaction of hepatitis C virus core protein with retinoid X receptor alpha modulates its transcriptional activity. Hepatology. 2002 Apr;35(4):937-46.

7. Jean-Christophe Meunier et al. Isolation and Characterization of Broadly Neutralizing Human Monoclonal Antibodies to the E1 Glycoprotein of Hepatitis C Virus. Journal of Virology. January 2008 vol. 82 no. 2 966-973.

8. Judith M. Gottwein et al. Novel Infectious cDNA Clones of Hepatitis C Virus Genotype 3a (Strain S52) and 4a (Strain ED43): Genetic Analyses and In Vivo Pathogenesis Studies. Journal of Virology. May 2010; 84(10): 5277-5293.

9. Hua Liang et al. Differential Effects of Hepatitis C Virus JFH1 on Human Myeloid and Plasmacytoid Dendritic Cells. Journal of Virology. Jun 2009; 83(11): 5693-5707.

10. Ryosuke Suzuki et al. Molecular Determinants for Subcellular Localization of Hepatitis C Virus Core Protein. Journal of Virology. Jan 2005; 79(2): 1271-1281.

11. Jiaren Sun et al. Hepatitis C Virus Core and Envelope Proteins Do Not Suppress the Host"s Ability To Clear a Hepatic Viral Infection. Journal of Virology. Dec 2001; 75(24): 11992-11998.

12. Anna R. Ciccaglione et al. Repression of Interferon Regulatory Factor 1 by Hepatitis C Virus Core Protein Results in Inhibition of Antiviral and Immunomodulatory Genes. Journal of Virology. Jan 2007; 81(1): 202-214.

13. Zhang-Xu Liu et al. Hepatitis C Virus Genotype 1b Core Protein Does Not Exert Immunomodulatory Effects on Virus-Induced Cellular Immunity. Journal of Virology. Feb 2002; 76(3): 990-997.

14. Jiaren Sun et al. Parenchymal Expression of CD86/B7.2 Contributes to Hepatitis C Virus-Related Liver Injury. Journal of Virology. Aug 2005; 79(16): 10730-10739.

15. Akito Sakai et al. In Vivo Study of the HC-TN Strain of Hepatitis C Virus Recovered from a Patient with Fulminant Hepatitis: RNA Transcripts of a Molecular Clone (pHC-TN) Are Infectious in Chimpanzees but Not in Huh7.5 Cells. Journal of Virology. Jul 2007; 81(13): 7208-7219.

16. Hiroki Kawamura et al. HCV core expression in hepatocytes protects against autoimmune liver injury and promotes liver regeneration in mice. Hepatology. Volume 44, Issue 4, pages 936–944, October 2006.

17. Tanja B. Jensen et al. Highly Efficient JFH1-Based Cell-Culture System for Hepatitis C Virus Genotype 5a: Failure of Homologous Neutralizing-Antibody Treatment to Control Infection. Journal of Infectious Diseases. (2008) 198 (12): 1756-1765.

18. Masaaki Korenaga et al. Hepatitis C Virus Core Protein Inhibits Mitochondrial Electron Transport and Increases Reactive Oxygen Species (ROS) Production. Nov, 2005, The Journal of Biological Chemistry, 280, 37481-37488.

19. Jones DM, Atoom AM, Zhang X, Kottilil S, Russell RS.A genetic interaction between the core and NS3 proteins of hepatitis C virus is essential for production of infectious virus. J Virol 2011; 23(85):12351-61.

20. Li YP, Ramirez S, Gottwein JM, Scheel TK, Mikkelsen L, Purcell RH, Bukh J.Robust full-length hepatitis C virus genotype 2a and 2b infectious cultures using mutations identified by a systematic approach applicable to patient strains. Proc Natl Acad Sci U S A 2012; 18(109):E1101-10.

21. Scheel TK, Prentoe J, Carlsen TH, Mikkelsen LS, Gottwein JM, Bukh J.Analysis of functional differences between hepatitis C virus NS5A of genotypes 1-7 in infectious cell culture systems. PLoS Pathog 2012; 5(8):e1002696.

22. Li YP, Ramirez S, Jensen SB, Purcell RH, Gottwein JM, Bukh J.Highly efficient full-length hepatitis C virus genotype 1 (strain TN) infectious culture system. Proc Natl Acad Sci U S A 2012; 48(109):19757-62.

23. Gottwein JM, Jensen SB, Li YP, Ghanem L, Scheel TK, Serre SB, Mikkelsen L, Bukh J. Combination treatment with hepatitis C virus protease and NS5A inhibitors is effective against recombinant genotype 1a, 2a, and 3a viruses. Antimicrob Agents Chemother 2013; 3(57):1291-303.

24. Scheel TK, Galli A, Li YP, Mikkelsen LS, Gottwein JM, Bukh J.Productive homologous and non-homologous recombination of hepatitis C virus in cell culture. PLoS Pathog 2013; 3(9):e1003228.

25. Galli A, Scheel TKH, Prentoe JC, Mikkelsen LS, Gottwein JM, Bukh J.Analysis of hepatitis C virus core/NS5A protein co-localization using novel cell culture systems expressing core-NS2 and NS5A of genotypes 1-7. J Gen Virol 2013; Pt 10(94):2221-35.

26. Holder KA, Stapleton SN, Gallant ME, Russell RS, Grant MD.Hepatitis C virus-infected cells downregulate NKp30 and inhibit ex vivo NK cell functions. J Immunol 2013; 6(191):3308-18.

27. Lyn RK, Singaravelu R, Kargman S, O"Hara S, Chan H, Oballa R, Huang Z, Jones DM, Ridsdale A, Russell RS, Partridge AW, Pezacki JP.Stearoyl-CoA desaturase inhibition blocks formation of hepatitis C virus-induced specialized membranes. Sci Rep 2014; (4):4549.

28. Saito K, Shirasago Y, Suzuki T, Aizaki H, Hanada K, Wakita T, Nishijima M, Fukasawa M.Targeting cellular squalene synthase, an enzyme essential for cholesterol biosynthesis, is a potential antiviral strategy against hepatitis C virus. J Virol 2015; 4(89):2220-32.

29. Tanida I, Shirasago Y, Suzuki R, Abe R, Wakita T, Hanada K, Fukasawa M.Inhibitory Effects of Caffeic Acid, a Coffee-Related Organic Acid, on the Propagation of Hepatitis C Virus. Jpn J Infect Dis 2015; 4(68):268-75.

30. Amador-Cañizares Y, Bernier A, Wilson JA, Sagan SM.miR-122 does not impact recognition of the HCV genome by innate sensors of RNA but rather protects the 5" end from the cellular pyrophosphatases, DOM3Z and DUSP11. Nucleic Acids Res 2018; 10(46):5139-5158.

31. Hung TC, Jassey A, Lin CJ, Liu CH, Lin CC, Yen MH, Lin LT.Methanolic Extract of Rhizoma Coptidis Inhibits the Early Viral Entry Steps of Hepatitis C Virus Infection. Viruses 2018; 12(10):.

32. Scheel TK, Gottwein JM, Carlsen TH, Li YP, Jensen TB, Spengler U, Weis N, Bukh J. Efficient culture adaptation of hepatitis C virus recombinants with genotype-specific core-NS2 by using previously identified mutations. J Virol 2011; 6(85):2891-906.

33. Kofahi HM, Taylor NGA, Hirasawa K, Grant MD, Russell RS.Hepatitis C Virus Infection of Cultured Human Hepatoma Cells Causes Apoptosis and Pyroptosis in Both Infected and Bystander Cells. Sci Rep 2016; (6):37433.