Preparation, Purification and Identification of Excreted/Secreted Antigens of Toxoplasma Gondii Tachyzoites

  S.H. Abdollahi¹ *

 �

 1- Assistant Professor, Department of Microbiology, Rafsanjan University of Medical Sciences, Rafsanjan, Iran

  Background: The Excreted/Secreted antigens (E/SA) of toxoplasma gondii tachyzoites are thought to play an important role in the pathogenesis of parasite and the hosts^, immunity against infection with this Parasite . These antigens are also useful for diagnosis and differentiation of acute from chronic phase of toxoplasmosis . Therefore the aim of the present study was to detect the best technique for preparation, purification and identification of these antigens.

  Materials and Methods: Tachyzoites (2×10⁸) were incubated at 37°C for 1hour under mild agitation in test–tubes containing 2ml of RPMI-1640 culture medium without FCS. After centrifugation for 15min at 1000g the E/SA containing supernatant were dialyzed and concentrated. E/SA were separated by gel filtration and ion exchange chromatography.

  Fractions of both chromatography columns were analyzed by Native–PAGE electropho_ resis.

  Results: E/SA were eluted in three and two Peaks by gel filtration and ion-exchange chromatography respectively. The second peak obtained from ion-exchange chromatograp_ hy produced one band in native – PAGE.

  Conclusion: The result of this experiment showed that ion-exchange chromatography under this conditions is a good tool for purification of E/SA .

  Keywords: Toxoplasma Gondii, Excreted/Secreted Antigens, Ion-exchange Choromatography, Electrophoresis

  * Corresponding author tel: (391) 5234003-5