Serving the Society Since 1986
高级搜索
Volume 27 Issue 3
June 2012

    Wen-juan Wang, Xiong Li, Li-hua Wang, Hu Shan, Lei Cao, Peng-cheng Yu, Qing Tang and Guo-dong Liang. Preparation and Identification of Anti-rabies Virus Monoclonal Antibodies*[J]. Virologica Sinica, 2012, 27(3): 172-178. doi: 10.1007/s12250-012-3242-0
    Citation: Wen-juan Wang, Xiong Li, Li-hua Wang, Hu Shan, Lei Cao, Peng-cheng Yu, Qing Tang, Guo-dong Liang. Preparation and Identification of Anti-rabies Virus Monoclonal Antibodies* .VIROLOGICA SINICA, 2012, 27(3) : 172-178.  http://dx.doi.org/10.1007/s12250-012-3242-0
    shu

    抗狂犬病病毒单克隆抗体的制备及鉴定

    cstr: 32224.14.s12250-012-3242-0
    • 1.

      青岛农业大学动物科技学院，青岛，266109，中国; 2中国疾病预防控制中心病毒病预防控制所，北京，102206，中国; 3新疆维吾尔族自治区疾病预防控制中心，新疆，830001，中国; 4中国科学院武汉病毒研究所，武汉，430071，中国

    • 通讯作者： 申辛欣, qtang04@sina.com; 
    • 收稿日期： 2012-02-08
      录用日期： 2012-03-28

    • 制备抗狂犬病病毒单克隆抗体，为研发快速特异的狂犬病病毒诊断方法奠定基础。以狂犬病病毒核蛋白、狂犬病病毒人用疫苗株（PV株）为免疫原免疫6-8周龄雌性BALB/c小鼠，取其脾细胞与SP2/0骨髓瘤细胞按常规方法进行融合；利用间接免疫荧光法筛选所获单抗细胞株；将获得特异稳定分泌抗狂犬病病毒单克隆抗体的杂交瘤细胞株经小鼠腹腔注射制备单抗腹水；对所得单抗进行了亚类及特异性和敏感性的系统鉴定。获得2株（3B12和4A12）特异稳定分泌抗狂犬病病毒单抗细胞株，其腹水效价分别为1：8000和1：10000，分泌单抗为抗狂犬病病毒IgG2a亚类。所得两株单抗细胞株能高效价稳定特异分泌抗狂犬病病毒单抗，且具有良好的稳定性和特异性，适宜应用于下一步狂犬病检测试剂的研发。

    Preparation and Identification of Anti-rabies Virus Monoclonal Antibodies*

    • 1.

      College of Animal Science and Veterinary Medicine, Qingdao Agricultural university, Qingdao 266109, China

    • 2.

      State Key Laboratory for Infectious Disease Prevention and Control, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China

    • 3.

      Xinjiang Center for Disease Control and Prevention, Xinjiang 830001, China

    • Corresponding author: Qing Tang, qtang04@sina.com
    • Received Date: 08 February 2012
      Accepted Date: 28 March 2012

      Fund Project: National Department Public Benefit Research Foundation 201103032Pathogens Network Monitoring Technology Research 2008ZX 10004-008

    • To provide a foundation for the development of rapid and specific methods for the diagnosis of rabies virus infection, anti-rabies virus monoclonal antibodies were prepared and rabies virus nucleoprotein and human rabies virus vaccine strain (PV strain) were used as immunogens to immunize 6-8 week old female BALB/c mice. Spleen cells and SP2/0 myeloma cells were fused according to conventional methods: the monoclonal cell strains obtained were selected using the indirect immunofluorescence test; this was followed by preparation of monoclonal antibody ascitic fluid; and finally, systematic identification of subclass, specificity and sensitivity was carried out. Two high potency and specific monoclonal antibodies against rabies virus were obtained and named 3B12 and 4A12, with ascitic fluid titers of 1:8000 and 1:10000, respectively. Both belonged to the IgG2a subclass. These strains secrete potent, stable and specific anti-rabies virus monoclonal antibodies, which makes them well suited for the development of rabies diagnosis reagents.
    • 加载中

      1. Berry J D, Jones S, Drebot M A, et al. 2004. Development and characterization of neutralizing monoclonal antibody to the SARS-coronavirus. J Virol Methods, 120: 67-96.

      2. Camila Z, Luan R P A, Paula P M, et al. 2011. Novel monoclonal antibodies that bind to wild and fixed rabies virus strains. J Virol Methods, 175: 66-73.
          doi: 10.1016/j.jviromet.2011.04.019

      3. Cao L, Tang Q, Tao X Y, et al. 2010. Identification and baculovirus expression of Rabies virus N gene, Chin J Exp Clin Virol, 5(26): 351-356. (in Chinese)

      4. de Kruif J, Bakker A B, Marissen W E, et al. 2007. A human monoclonal antibodies cocktail as a novel component of rabies postexposure prophylaxis. Annu Rev Med, 58: 359-368.
          doi: 10.1146/annurev.med.58.061705.145053

      5. Harlow E, Lane D. 1988. In: Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory, p219-220.

      6. Kohler G, Milstein C. 1975. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, 256(5517): 495-497.
          doi: 10.1038/256495a0

      7. Lembo T, Niezgoda M, Velasco-Villa A, et al. 2006. Evaluation of a direct, rapid immunohisto chemical test for rabies diagnosis. Emerg Infect Dis, 12:310-313.

      8. Liu W T, Yu P P, Li Z H. 2011. Purification and Application of the Rabies Viral Monoclonal Antibody. Chin J Vet Drug. 45(4):5-8.(in Chinese)

      9. Meslin F X, Kaplan M M, koprowski H. 1996. Laboratory techniques in rabies. Geneva: World Health Organization, p133-134.

      10. Muhamuda K, Madhusudana S N, Ravi V. 2007. Use of neutralizing murine monoclonal antibodies to rabies glycoprotein in passive immunotherapy against rabies. Hum Vaccine, 3(5): 192-195.
          doi: 10.4161/hv.3.5.4386

      11. Wiktor T J. 1978. Monoclonal antibodies against rabies virus produced by somatic cell hybridization: Detection of antigenic variants. Immunology, 75: 3938-3942.

      12. Yu P C, Lv X J, Shen X X, et al. 2010. The establishment of a rapid fluorescent focus inhibition test for testing rabies virus neutralizing antibody. Chin J Epidemiol, 31(4): 438-441. (in Chinese)

      13. Zhang S F, Liu Y, Zhang F, et al. 2009. Competitive ELISA using a rabies glycoprotein-transformed cell line to semi-quantify rabies neutralizing-related antibodies in dogs. Vaccine, 27:2108-2113. (in Chinese)
          doi: 10.1016/j.vaccine.2009.01.126

      14. Zhou Y, Li Y S, Pan F G, et al. 2006. The study of purified methods of mice ascitic fluid IgG McAb. Heilongjiang Anim Sci Vet Med, 10:14-16. (in Chinese)

    • 加载中

    Figures(2) / Tables(4)

    PDF

    Article Metrics

    Article views(7820) PDF downloads(19) Cited by(0)

    Related
    Proportional views
      通讯作者: 陈斌, bchen63@163.com
      • 1. 

        沈阳化工大学材料科学与工程学院 沈阳 110142

      1. 本站搜索
      2. 百度学术搜索
      3. 万方数据库搜索
      4. CNKI搜索

      Preparation and Identification of Anti-rabies Virus Monoclonal Antibodies*

        Corresponding author: Qing Tang, qtang04@sina.com
      • 1. College of Animal Science and Veterinary Medicine, Qingdao Agricultural university, Qingdao 266109, China
      • 2. State Key Laboratory for Infectious Disease Prevention and Control, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
      • 3. Xinjiang Center for Disease Control and Prevention, Xinjiang 830001, China
      • Received Date: 08 February 2012
      • Accepted Date: 28 March 2012
      Fund Project:  National Department Public Benefit Research Foundation 201103032Pathogens Network Monitoring Technology Research 2008ZX 10004-008

      Abstract: To provide a foundation for the development of rapid and specific methods for the diagnosis of rabies virus infection, anti-rabies virus monoclonal antibodies were prepared and rabies virus nucleoprotein and human rabies virus vaccine strain (PV strain) were used as immunogens to immunize 6-8 week old female BALB/c mice. Spleen cells and SP2/0 myeloma cells were fused according to conventional methods: the monoclonal cell strains obtained were selected using the indirect immunofluorescence test; this was followed by preparation of monoclonal antibody ascitic fluid; and finally, systematic identification of subclass, specificity and sensitivity was carried out. Two high potency and specific monoclonal antibodies against rabies virus were obtained and named 3B12 and 4A12, with ascitic fluid titers of 1:8000 and 1:10000, respectively. Both belonged to the IgG2a subclass. These strains secrete potent, stable and specific anti-rabies virus monoclonal antibodies, which makes them well suited for the development of rabies diagnosis reagents.

        HTML