Skip to main content

Advertisement

SpringerLink
Log in

Production of monoclonal antibodies and development of an antigen capture ELISA directed against the envelope glycoprotein GP of Ebola virus

  • Original Investigation
  • Published:
Medical Microbiology and Immunology Aims and scope Submit manuscript

Abstract

Ebola virus (EBOV) causes severe outbreaks of Ebola hemorrhagic fever in endemic regions of Africa and is considered to be of impact for other parts of the world as an imported viral disease. To develop a new diagnostic test, monoclonal antibodies to EBOV were produced from mice immunized with inactivated EBOV species Zaire. Antibodies directed against the viral glycoprotein GP were characterized by ELISA, Western blot and immunofluorescence analyses. An antigen capture ELISA was established, which is specific for EBOV-Zaire and shows a sensitivity of approximately 103 plaque-forming units/ml. Since the ELISA is able to detect even SDS-inactivated EBOV in spiked human sera, it could complement the existing diagnostic tools in the field and in routine laboratories where high containment facilities are not available.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4.

Similar content being viewed by others

References

  1. Baize S, Leroy EM, Courbot MC, Capron M, Landsoud-Soukate J, Debré P, Fisher-Hoch SP, McCormick JB, Georges AJ (1999) Defective humoral responses and extensive intravascular apoptosis are associated with fatal outcome in Ebola virus-infected patients. Nat Med 5:423–426

    Article  PubMed  Google Scholar 

  2. Becker S, Feldmann H, Will C, Slenczka W (1992) Evidence of occurrence of filoviruses antibodies in humans and imported monkeys: do subclinical filovirus infections occur worldwide? Med Microbiol Immunol 181:43–55

    PubMed  Google Scholar 

  3. Becker S, Klenk HD, Muhlberger E (1996) Intracellular transport and processing of the Marburg virus surface protein in vertebrate and insect cells. Virology 225:145–55

    Article  PubMed  Google Scholar 

  4. Becker S, Rinne C, Hofsass U, Klenk HD, Muhlberger E (1998) Interactions of Marburg virus nucleocapsid proteins. Virology 249:406–417

    Article  PubMed  Google Scholar 

  5. Borisevich IV, Mikhailov VV, Potryvaeva NV, Malinkin IN, Kirillov AP, Krasnianskii VP, Markov VI, Makhlai AA, Lebedinskaia EV (1996) Development of the immunoenzyme test-system for detection of Ebola virus antigen. Vopr Virus 41:232–234

    PubMed  Google Scholar 

  6. Bowen ETW, Platt GS, Lloyd G, Baskerville A, Harris WJ, Vella EE (1977) Viral haemorrhagic fever in southern Sudan and northern Zaire. Lancet I:571–572

    Article  Google Scholar 

  7. Centers for Disease Control (1989) Ebola virus infection in imported primates—Virginia 1989. MMWR Morb Mortal Wkly Rep 38:831–832

    PubMed  Google Scholar 

  8. Centers for Disease Control and Prevention (1996) Ebola-Reston virus infection among quarantined primates—Texas 1996. MMWR Morb Mortal Wkly Rep 45:314–316

    PubMed  Google Scholar 

  9. Edmond RTD, Evans B, Bowen ETW, Lloyd G (1977) A case of Ebola virus infection. Br Med J 2:541–544

    PubMed  Google Scholar 

  10. Feldmann H, Slenczka W, Klenk HD (1996) Classification, structure, and replication of filoviruses. Curr Top Microbiol Immunol 236:1–22

    Google Scholar 

  11. Feldmann H, Volchkov VE, Volchkova VA, Stroher U, Klenk HD (2001) Biosynthesis and role of filovirus glycoproteins. J Gen Virol 82:2839–2848

    PubMed  Google Scholar 

  12. Felgner JH, Kumar R, Sridhar CN, Wheeler CJ, Tsai YJ, Border R, Ramsey P, Martin M, Felgner PL (1994) Enhanced gene delivery and mechanism studies with a novel series of cationic lipid formulations. J Biol Chem 269:2550–2561

    PubMed  Google Scholar 

  13. Geisbert TW, Jahrling PB (1995) Differentiation of filoviruses by electron microscopy. Virus Res 39:129–150

    Article  PubMed  Google Scholar 

  14. Grunow R, Giese R, Porstmann T, Doepel H, Haensel K, Bähr R von (1990) Development and biological testing of human and murine monoclonal antibodies against HIV antigens. Z Klin Med 45:367–369

    Google Scholar 

  15. Ivanoff B, Duquesnoy P, Languillat G, Saluzzo JF, Georges A, Gonzalez JP, McCormick J (1982) Haemorrhagic fever in Gabon. I. Incidence of Lassa, Ebola and Marburg viruses in Haut-Ogooué. Trans R Soc Trop Med Hyg 76:719–720

    PubMed  Google Scholar 

  16. Jahrling PB, Geisbert TW, Dalgard DW, Johnson ED, Ksiazek TG, Hall WC, Peters CJ (1990) Preliminary report: isolation of Ebola virus from monkeys imported to USA. Lancet 335:502–505

    PubMed  Google Scholar 

  17. Johnson BK, Wambui C, Ocheng D, Gichogo A, Oogo S, Libondo D, Gitau LG, Tukei PM, Johnson ED (1986) Seasonal variation in antibodies against Ebola virus in Kenyan fever patients. Lancet I:1160

    Article  Google Scholar 

  18. Johnson KM, Webb PA, Lange JV, Murphy FA (1977) Isolation and partial characterization of a new virus causing acute haemorrhagic fever in Zaire. Lancet I:569–571

    Article  Google Scholar 

  19. Johnson KM, Elliott LH, Heymann DL (1981) Preparation of polyvalent viral immunofluorescent intracellular antigens and use in human serosurveys. J Clin Microbiol 14:527–529

    PubMed  Google Scholar 

  20. Kearney JF, Radbruch A, Liesegang B, Rajewsky K (1979) A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immun 123:1548–1550

    CAS  PubMed  Google Scholar 

  21. Kiley MP, Bowen ETW, Eddy GA, Isaacson M, Johnson KM, McCormick JB, Murphy FA, Pattyn SR, Peters D, Prozesky OW, Regnery RL, Simpson DIH, Slenczka W, Sureau P, van der Groen G, Webb PA, Wulff H (1982) Filoviridae: a taxonomic home for Marburg and Ebola virus? Intervirology 18:24–32

    PubMed  Google Scholar 

  22. Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497

    PubMed  Google Scholar 

  23. Ksiazek TG, Rollin PE, Jahrling PB, Johnson E, Dalgard DW, Peters CJ (1992) Enzyme immunosorbent assay for Ebola virus antigens in tissues of infected primates. J Clin Microbiol 30:947–950

    PubMed  Google Scholar 

  24. Ksiazek TG, Rollin PE, Williams AJ, Bressler DS, Martin ML, Swanepoel R, Burt FJ, Leman PA, Khan AS, Rowe AK, Mukunu R, Sanchez A, Peters CJ (1999) Clinical virology of Ebola hemorrhagic fever (EHF): virus, virus antigen, and IgG and IgM antibody findings among EHF patients in Kikwit, Democratic Republic of the Congo, 1995. J Infect Dis 179 [Suppl 1]:177–187

  25. LeGuenno B, Formentry P, Wyers M, Gounon P, Walker F, Boesch C (1995) Isolation and partial characterization of a new strain of Ebola virus. Lancet 345:1271–1274

    Article  PubMed  Google Scholar 

  26. Lenz O, Klenk HD, Slenczka W, Feldmann H (1998) Infektionen durch Marburg- und Ebolavirus. Epidem Bull 45:317–319

    Google Scholar 

  27. Leroy EM, Baize S, Lu CY, McCormick JB, Georges AJ, Georges-Courbot MC, Lansoud-Soukate J, Fisher-Hoch SP (2000) Diagnosis of Ebola haemorrhagic fever by RT-PCR in an epidemic setting. J Med Virol 60:463–467

    Article  PubMed  Google Scholar 

  28. Lucht A, Grunow R, Möller P, Feldmann H, Becker S (2003). Development, characterization and use of monoclonal VP40-antibodies for the detection of Ebola virus. J Virol Methods 111:21–28

    Article  PubMed  Google Scholar 

  29. Maryama T, Rodriguez LL, Jahrling PB, Sanchez A, Khan AS, Nichol ST, Peters CJ, Parren PW, Burton DR (1999). Ebola virus can be effectively neutralized by antibody produced in natural human infection. J Virol 73:6024–6030

    PubMed  Google Scholar 

  30. Merzlikin NV, Chepurnov AA, Istomina NN, Ofitserviv VI, Vorobéva MS (1995) Development and application of an immunoenzyme test system for diagnosing Ebola fever. Vopr Virus 40:31–35

    PubMed  Google Scholar 

  31. Mühlberger E, Sanchez A, Randolf A, Will C, Kiley MP, Klenk HD, Feldmann H (1992) The nucleotide sequence of the L gene of Marburg virus, a filovirus, homologies with paramyxoviruses and rhabdoviruses. Virology 187:534–537

    PubMed  Google Scholar 

  32. Nakane PK, Kawaoi A (1974) Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem 22:1084–1091

    CAS  PubMed  Google Scholar 

  33. Niikura M, Ikegami T, Saijo M, Kurane I, Miranda ME, Morikawa S (2001) Detection of Ebola viral antigen by enzyme-linked immunosorbentassay using a novel monoclonal antibody to nucleoprotein. J Clin Microbiol 39:3267–3271

    Article  PubMed  Google Scholar 

  34. Okome-Nkoumou M, Kombila M (1999) A case of Ebola haemorrhagic fever in Libreville, fatal after evacuation to South Africa. Med Trop 59:411

    CAS  Google Scholar 

  35. Pattyn SR, Jacob W, Van Der Groen G, Piot P (1977) Isolation of Marburg-like virus from a case of haemorrhagic fever in Zaire. Lancet I:573–574

    Article  Google Scholar 

  36. Rollin PE, Ksiazek TG, Jahrling PB, Haines M, Peters CJ (1990) Detection of Ebola-like viruses by immunofluorescence. Lancet 336:1591

    PubMed  Google Scholar 

  37. Rowe AK, Bertolli J, Khan AS, Mukum R, Muyembe-Tamfun JJ, Bressler D, Williams AJ, Peters CJ, Rodriguez L, Feldmann H, Nichol ST, Rollin PE, Ksiazek TG (1999) Clinical, virologic, and immunologic follow up of convalescent Ebola haemorrhagic fever patients and their household contacts, Kikwit, Democratic Republic of the Congo. J Infect Dis 179 Suppl 1:S28-S35

    Google Scholar 

  38. Sadek RF, Khan AS, Stevens G, Peters CJ, Ksiazek TG (1999) Ebola haemorrhagic fever, Democratic Republic of the Congo, 1995: Determinants of survival. J Infect Dis 179 [Suppl 1]:S24–S27

  39. Sanchez A, Feldmann H (1996) Detection of Marburg and Ebola virus infections by polymerase chain reaction assays. In: Becker Y (ed) Frontiers of Virology—Diagnosis of human viruses by polymerase chain reaction technology. Springer, Berlin Heidelberg New York, pp 411–418

  40. Sanchez A, Trappier SG, Mahy BWJ, Peters CJ, Nichol ST (1996) The virion glycoproteins of Ebola viruses are encoded in two reading frames and are expressed through descriptional editing. Proc Natl Acad Sci USA 93:3602–3607

    CAS  PubMed  Google Scholar 

  41. Slenczka W, Shu HL, Piepenburg G, Siegert R. (1968) “Marburg virus” antigen demonstration by immunofluorescence in organs of infected guinea pigs. Dtsch Med Wochenschr 93:612–616

    PubMed  Google Scholar 

  42. Slenczka W, Rietschel M (1984) Seroepidemiologische Untersuchungen über das Vorkommen von Antikörpern gegen Marburg- und Ebola-Virus. Mitt Oest Ges Tropenmed Parasitol 6:53–60

    Google Scholar 

  43. Sutter G, Ohlmann M, Erfle V (1995) Non-replicating vaccinia vector efficiently expresses bacteriophage T7 RNA polymerase. FEBS Lett 371:9–12

    Article  PubMed  Google Scholar 

  44. Van der Waals FW, Pomeroy KL, Goudsmit J, Asher DM, Gajdusek DC (1986) Hemorrhagic fever virus infections in an isolated rainforest area of central Liberia. Limitations of the indirect immunofluorescence slidetest for antibody screening in Africa. Trop Geogr Med 38:209–214

    PubMed  Google Scholar 

  45. Van Regenmortel MHV, Fauquet CM, Bishop DHL, Carstens EB, Estes MK, Lemon SM, Maniloff J, Mayo MA, McGeoch DJ, Pringle CR, Wickner RB (2000) Virus taxonomy: The classification and nomenclature of viruses. The Seventh Report of the International Committee on Tasonomy of viruses (Book). Virus Taxonomy, VIIth report of ICTV. Academic Press, San Diego, p 1167

    Google Scholar 

  46. Volchkov VE, Becker S, Volchkova VA, Ternovoj VA, Kotov AN, Netesov SV, Klenk HD (1995) GP mRNA of Ebola virus is edited by the Ebola virus polymerase and by T7 and vaccinia virus polymerases. Virology 214:421–430

    Article  PubMed  Google Scholar 

  47. Volchkov VE, Feldmann H, Volchkva VA, Klenk HD (1998) Processing of the Ebola virus glycoprotein by the proprotein convertase furin. Proc Nat Acad Sci USA 95:5762–5767

    Article  PubMed  Google Scholar 

  48. Volchkov VE,Volchkova VA, Slenczka W, Klenk HD, Feldmann H (1998) Release of viral glycoproteins during Ebola virus infection. Virology 245:110–119

    Article  PubMed  Google Scholar 

  49. Wilson MB, Nakane PK (1976) The covalent coupling of proteins to periodate-oxidized sephadex: a new a approach to immunoadsorbent preparation. J Immunol Methods 12:171–181

    Article  PubMed  Google Scholar 

  50. World Health Organization (1976) Suspected viral haemorrhagic fever outbreaks in Sudan and Zaire. Wkly Epidemiol Rec 51:321

    Google Scholar 

  51. World Health Organization (1992) Viral haemorrhagic fever in imported monkeys. Wkly Epidemiol Rec 67:142–143

    PubMed  Google Scholar 

  52. Zaki SR, Shieh WJ, Greer PW, Goldsmith CS, Ferebee T, Katshitshi J, Tshioko FK, Bwaka MA, Swanepoel R, Calain P, Khan AS, Lloyd E, Rollin PE, Ksiazek TG, Peters CJ (1999) A novel histochemical assay for the detection of Ebola virus in skin: implications for diagnosis, spread and surveillance of Ebola hemorrhagic fever. Commission de Lutte contre les Epidemies a Kikwit. J Infect Dis 179 [Suppl 1]:36–47

Download references

Acknowledgements

This work was supported by the Bundesministerium der Verteidigung (Sonderforschungsauftrag 23Z1-S-439902) and by the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 286, TP A6 and Sonderforschungsbereich 535 TP A4 and B9) and the Canadian Institutes of Health Research (MOP-43921). The authors wish to thank Viktor Volchkov for helpful discussion. Thanks to N. Romhart and E. Zeman for technical assistance.

Author information

Author notes

  1. Christian Otterbein

    Present address: Authority of the district of Rosenheim, Public Veterinary Office, Am Klafferer 3, , 83043, Bad Aibling, Germany

  2. Heinz Feldmann

    Present address: Special Pathogens Program, National Microbiology Laboratory, Health Canada, Canadian Science Centre for Human and Animal Health, Winnipeg, Canada

Authors and Affiliations

  1. Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, 80937, Munich, Germany

    Andreas Lucht, Roland Grunow & Christian Otterbein

  2. Institute of Virology, Philipps-University Marburg, Robert-Koch-Strasse 17, , 35037, Marburg, Germany

    Peggy Möller, Heinz Feldmann & Stephan Becker

Authors
  1. Andreas Lucht
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Roland Grunow
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Christian Otterbein
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Peggy Möller
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Heinz Feldmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Stephan Becker
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Andreas Lucht.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lucht, A., Grunow, R., Otterbein, C. et al. Production of monoclonal antibodies and development of an antigen capture ELISA directed against the envelope glycoprotein GP of Ebola virus. Med Microbiol Immunol 193, 181–187 (2004). https://doi.org/10.1007/s00430-003-0204-z

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00430-003-0204-z

Keywords

Search

Navigation