Production of a monoclonal antibody specific for Citrus Tristeza Virus


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Ozturk S., Cirakoglu B.

FOOD AND AGRICULTURAL IMMUNOLOGY, vol.15, no.1, pp.65-73, 2003 (Journal Indexed in SCI) identifier identifier

  • Publication Type: Article / Article
  • Volume: 15 Issue: 1
  • Publication Date: 2003
  • Doi Number: 10.1080/0954010031000138113
  • Title of Journal : FOOD AND AGRICULTURAL IMMUNOLOGY
  • Page Numbers: pp.65-73

Abstract

In this work Citrus Tristeza Virus (CTV) was purified from the bark and midrib tissue of infected plants by pulverization of frozen tissue samples in a mortar with extraction buffer, with polyethylene glycol (PEG), and by sucrose gradient centrifugation. In an alternative purification method, CTV was purified by Sephacryl 300 column chromatography after precipitation by PEG. A hybridoma line secreting monoclonal antibody (MAb) highly specific for the CTV was generated by fusion of splenic lymphocytes of immunized BALB/c mice with non-secreting mouse myeloma cells followed by selection in hypoxanthine-aminopterin-thymidine (HAT) medium, and finally, by screening culture fluids for CTV affinity in ELISA. The MAb produced by the CTV-IG8 hybridoma clone was highly specific to CTV and did not react with non-infected citrus plant extracts.