Current Microbiology

, Volume 21, Issue 3, pp 175–180 | Cite as

Purification of the bacterium-like organism associated with greening disease of citrus by immunoaffinity chromatography and monoclonal antibodies

  • Sandrine Villechanoux
  • Monique Garnier
  • Joseph M. Bové


An immunoaffinity chromatographic procedure with monoclonal antibodies (MA) has been developed for purification of the uncultivable, bacterium-like organism associated with greening disease of citrus. The greening organism (GO) was partially purified from leaf midribs of infected periwinkle plants by differential centrifugation. The GO present in such preparations was retained on an affinity matrix consisting of CNBr-activated Sepharose 4B on which GO-specific MA had been covalently linked. The unbound plant material was washed from the matrix, and the GOs were eluted with 0.1M glycine (pH 11.5). Purified GOs were compared with organisms observed in the initial plant preparation by both immunofluorescence and electron microscopic techniques. The morphology and serological characteristics of the GO were retained following purification procedures.


Monoclonal Antibody Purification Procedure Microscopic Technique Differential Centrifugation Chromatographic Procedure 
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Copyright information

© Springer-Verlag New York Inc. 1990

Authors and Affiliations

  • Sandrine Villechanoux
    • 1
  • Monique Garnier
    • 1
  • Joseph M. Bové
    • 1
  1. 1.Laboratoire de biologie cellulaire et moléculaireINRA et université de bordeauxPont de la MayeFrance

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