The rabies virus is a neurotropic virus that causes fatal disease in humans and animals. However, not all commercial antibodies against rabies virus (RABV) structural proteins are generally available, and production of high-quality monoclonal antibodies (MAbs) requires high purification of virus particles and special facilities and is time-consuming. By using RABV-infected suckling mouse brain as antigens in this study, 11 hybridoma cells secreting MAbs against RABV were obtained, which showed strong reactivity with RABV-infected Vero cells in immunofluorescence assay. Among the 11 MAbs, three MAbs (1B11, 1C8, and 8H12) showed a neutralizing effect to RABV, while MAb 4B7 recognized the recombinant nucleoprotein (N) of RABV expressed in Vero cells; seven MAbs (1H3, 3H7, 4E7, 4G3, 5A10, 6C9, and 7B3) reacted specifically with phosphoprotein (P) of RABV. The MAbs developed in this study will be useful in establishing a diagnostic test and study on the interactions between RABV and its host.
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References
1.
WarnerC, FekaduM, WhitfieldS, and ShaddockJ: Use of anti-glycoprotein monoclonal antibodies to characterize rabies virus in formalin-fixed tissues. J Virol Methods, 1999; 77:69–74.
2.
ZhaoXL, YinJ, ChenWQ, JiangM, YangG, and YangZH: Generation and characterization of human monoclonal antibodies to G5, a linear neutralization epitope on glycoprotein of rabies virus, by phage display technology. Microbiol Immunol, 2008; 52:89–93.
3.
WiktorTJ, and KoprowskiH: Monoclonal antibodies against rabies virus produced by somatic cell hybridization: detection of antigenic variants. Proc Natl Acad Sci USA, 1978; 75:3938–3942.
4.
Nadin-DavisSA, Fehlner-GardinerC, SheenM, and WandelerAI: Characterization of a panel of anti-phosphoprotein monoclonal antibodies generated against the raccoon strain of rabies virus. Virus Res, 2010; 152:126–136.
5.
ZanlucaC, AiresLRD, MuellerPP, dos SantosVV, CarrieriML, PintoAR, and ZanettiCR: Novel monoclonal antibodies that bind to wild and fixed rabies virus strains. J Virol Methods, 2011; 175:66–73.
6.
ChampionJM, KeanRB, RupprechtCE, NotkinsAL, KoprowskiH, DietzscholdB, and HooperDC: The development of monoclonal human rabies virus-neutralizing antibodies as a substitute for pooled human immune globulin in the prophylactic treatment of rabies virus exposure. J Immunol Methods, 2000; 235:81–90.
7.
DorfmanN, DietzscholdB, KajiyamaW, FuZF, KoprowskiH, and NotkinsAL: Development of human monoclonal-antibodies to rabies. Hybridoma, 1994; 13:397–402.
8.
LafonM, EdelmanL, BouvetJP, LafageM, and MontchatreE: Human monoclonal-antibodies specific for the rabies virus glycoprotein and N protein. J Gen Virol, 1990; 71:1689–1696.
9.
WangWJ, LiX, WangLH, ShanH, CaoL, YuPC, TangQ, and LiangGD: Preparation and identification of anti-rabies virus monoclonal antibodies. Virol Sinica, 2012; 27:172–178.
10.
TurkiI, HammamiA, KharmachiH, and MousliM: Engineering of a recombinant trivalent single-chain variable fragment antibody directed against rabies virus glycoprotein G with improved neutralizing potency. Mol Immunol, 2013; 57:66–73.
11.
Cai YQMYP, ShenHG, GuoJQ, and ZhouJY: High level expression of nucleoprotein gene of rabies virus in Escherichia coli. Vet Sci China, 2007; 37:145–149.
12.
KöhlerG, and MilsteinC: Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, 1975; 256:495–497.
13.
KhawplodP, InoueK, ShojiY, WildeH, UbolS, NishizonoA, KuraneI, and MorimotoK: A novel rapid fluorescent focus inhibition test for rabies virus using a recombinant rabies virus visualizing a green fluorescent protein. J Virol Methods, 2005; 125:35–40.
14.
ZhangJ, WuX, ZanJ, WuY, YeC, RuanX, and ZhouJ: Cellular chaperonin CCTgamma contributes to rabies virus replication during infection. J Virol, 2013; 87:7608–7621.
15.
SmithJS, YagerPA, and BaerGM: A rapid fluorescent focus inhibition test (RFFIT) for determining rabies virus-neutralizing antibody. In: Laboratory Techniques in Rabies, 4th ed.MeslinF-X, KaplanMM, and KoprowskiH, Eds. World Health Organization: Geneva, 1996; 181–191.
16.
ZhangJ, LiP, ZhangW, ZhangQ, DingX, ChenX, WuW, and ZhangX: Production and characterization of monoclonal antibodies against aflatoxin g(1). Hybridoma, 2009; 28:67–70.
17.
ZhangX, MaG, LiY, JiangX, HeJ, and ZhouJ: Characterization of monoclonal antibody against replication-associated protein of porcine circovirus. DNA Cell Biol, 2009; 28:23–29.
18.
ZhengX, HongL, LiY, GuoJ, ZhangG, and ZhouJ: In vitro expression and monoclonal antibody of RNA-dependent RNA polymerase for infectious bursal disease virus. DNA Cell Biol, 2006; 25:646–653.
19.
ZhengX, HongL, ShiL, GuoJ, SunZ, and ZhouJ: Proteomics analysis of host cells infected with infectious bursal disease virus. Mol Cell Proteomics, 2008; 7:612–625.
20.
ZhouJY, ShangSB, GongH, ChenQX, WuJX, ShenHG, ChenTF, and GuoJQ: In vitro expression, monoclonal antibody and bioactivity for capsid protein of porcine circovirus type II without nuclear localization signal. J Biotechnol, 2005; 118:201–211.
21.
BlondelD, LahayeX, VidyA, PomierC, ObiangL, HarperF, and GaudinY: Functional characterization of Negri bodies (nbs) in rabies virus-infected cells: evidence that NBS are sites of viral transcription and replication. J Virol, 2009; 83:7948–7958.
22.
PataS, TayapiwatanaC, and KasinrerkW: Three different immunogen preparation strategies for production of CD4 monoclonal antibodies. Hybridoma, 2009; 28:159–165.
23.
AstoulE, LafageM, and LafonM: Rabies superantigen as a Vbeta T-dependent adjuvant. J Exp Med, 1996; 183:1623–1631.
