Summary
It has been claimed recently [Dietzgen (1986) Arch Virol 91: 163–173] that a series of monoclonal antibodies (Mabs) produced against the nepovirus, arabis mosaic virus (ArMV) cross-reacted with the tobamovirus, tobacco mosaic virus (TMV). In the present report, this alleged cross-reactivity was re-examined by two ELISA procedures using Mabs produced against each of the two viruses. It was found that when highly concentrated preparations of Mabs were used, all antibodies reacted in a nonspecific manner with several plant viruses. However, when defatted milk instead of bovine serum albumin was used both as blocking agent and as diluent for the Mabs, the spurious cross-reactions between unrelated viruses were abolished. The use of milk as blocking agent did not prevent the detection of genuine cross-reactions between related nepoviruses.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Al Moudallal Z, Briand JP, Van Regenmortel MHV (1982) Monoclonal antibodies as probes of the antigenic structure of tobacco mosaic virus. EMBO J 1: 1005–1010
Al Moudallal Z, Altschuh D, Briand JP, Van Regenmortel MHV (1984) Comparative sensitivity of different ELISA procedures for detecting monoclonal antibodies. J Immunol Methods 58: 35–43
Al Moudallal Z, Briand JP, Van Regenmortel MHV (1985) A major part of the polypeptide chain of tobacco mosaic virus protein is antigenic. EMBO J 4: 1231–1235
Altschuh D, Van Regenmortel MHV (1983) Refutation by ELISA of the alleged antigenic relationship between an isometric and a rod-shaped plant virus. Int J Microbiol 1: 13–17
Bercks R, Querfurth G (1971) Serologische Beziehungen zwischen einem gestreckten (tobacco mosaic) und einem isometrischen (cocksfoot mild mosaic) Virus. Phytopathol Z 90: 223–243
Crowle AJ (1973) Immunodiffusion, 2nd edn. Academic Press, New York
Dietzgen RG (1983) Monoklonale Antikörper gegen Pflanzenviren: Herstellung, Reinigung und Charakterisierung. Doctoral dissertation, University of Tübingen, Federal Republic of Germany
Dietzgen RG (1986) Immunological properties and biological function of monoclonal antibodies to tobacco mosaic virus. Arch Virol 87: 73–86
Dietzgen RG (1986) Characterisation of antigenic structure of arabic mosaic virus with monoclonal antibodies. Arch Virol 91: 163–173
Dietzgen RG, Francki RIB (1987) Nonspecific binding of immunoglobulins to coat proteins of certain plant viruses in immunoblots and indirect ELISA. J Virol Methods 15: 159–164
Dietzgen RG, Zaitlin M (1986) Tobacco mosaic virus coat protein and the large subunit of the host protein ribulose 1,5-biphosphate carboxylase share a common antigenic determinant. Virology 155: 262–266
Dore I, Weiss E, Altschuh D, Van Regenmortel MHV (1988) Visualization by electron microscopy of the location of tobacco mosaic virus epitopes reacting with monoclonal antibodies in enzyme immunoassay. Virology 162: 279–289
Huss B (1986) Diagnostic des virus du Court-Noué de la vigne et études d'interactions entre isolats: utilisation d'anticorps monoclonaux. Doctoral dissertation, University of Orsay, France
Huss B, Muller S, Sommermeyer G, Walter B, Van Regenmortel MHV (1987) Grapevine fanleaf virus monoclonal antibodies: their use to distinguish different isolates. J Phytopathol 119: 358–370
Koenig R, Burgermeister W (1986) Applications of immuno-blotting in plant virus diagnosis. In: Jones RAC, Torrance L (eds) Developments in applied biology I. Developments and applications in virus testing. Proceedings of a conference at the University of Cambridge, April 1985. Association of Applied Biologists, Wellesbourne, UK, pp 121–137
Shepard JF, Secor GA, Purcifull DE (1974) Immunochemical cross-reactivity between the dissociated capsid proteins of PVY group plant viruses. Virology 58: 464–475
Sluyser M, Rümke Ph, Hekman A (1969) Antigenicity of histones: comparative studies on histones with very high lysine content from various sources. Immunochemistry 6: 494–498
Sotirov N, Johns EW (1972) Rabbit precipitin antibody to chicken erythrocyte histone F2C. J Immunol 109: 686–691
Stollar BD, Rezuke W (1978) Separation of anti-histone antibodies from nonimmune histone-precipitating serum proteins, predominantly α2-macroglobulin. Arch Biochem Biophys 190: 398–404
Van Regenmortel MHV (1981) Tobamoviruses. In: Kurstak E (ed) Handbook of plant virus infections and comparative diagnosis. Elsevier/North Holland Biochemical Press, Amsterdam, pp 541–564
Van Regenmortel MHV (1982) Serology and immunochemistry of plant viruses. Academic Press, New York
Van Regenmortel MHV, Burckard J (1980) Detection of a wide spectrum of tobacco mosaic virus strains by indirect enzyme-linked immunosorbent assays (ELISA). Virology 106: 327–334
Vogt RF Jr, Phillips DL, Henderson LO, Whitfield W, Spierto FW (1987) Quantitative differences among various proteins as blocking agents for ELISA microtiter plates. J Immunol Methods 101: 43–50
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Zimmermann, D., Van Regenmortel, M.H.V. Spurious cross-reactions between plant viruses and monoclonal antibodies can be overcome by saturating ELISA plates with milk proteins. Archives of Virology 106, 15–22 (1989). https://doi.org/10.1007/BF01311034
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01311034