Production of a monoclonal antibody specific for Citrus Tristeza Virus


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Ozturk S., Cirakoglu B.

FOOD AND AGRICULTURAL IMMUNOLOGY, cilt.15, ss.65-73, 2003 (SCI İndekslerine Giren Dergi) identifier identifier

  • Cilt numarası: 15 Konu: 1
  • Basım Tarihi: 2003
  • Doi Numarası: 10.1080/0954010031000138113
  • Dergi Adı: FOOD AND AGRICULTURAL IMMUNOLOGY
  • Sayfa Sayıları: ss.65-73

Özet

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.