ÖZLEM BÜYÜKTANIR YAŞ, Tamer Yağcı, ARZU FINDIK, TUBA YILDIRIM, Nevzat YURDUSEV

Generation of mouse hybridomas secreting anti-Salmonella Enteritidis antibodies and their preliminary characterization

Salmonella Enteritidis’e karşı monoklonal antikor üretmek amacıyla BALB/c fareler inaktif bakteri ile periton içi bağışıklandı. Beşinci bağışıklamada en yüksek yanıtı veren hayvana ait dalak hücreleri fare Sp2/0 miyeloma hücrelerinin füzyon partneri olarak kullanıldı. IgM ve IgG izotipi antikorlar salgılayan toplam 6 adet kararlı hibridoma hücresi elde edildi. Bu hibridomaların Western blot analizinde S. Enteritidis’e ait 73, 59 ve 42 kDa relatif molekül ağırlıklarına sahip 3 antijen ile reaksiyon verdiği belirlendi. 59 kDa molekül Salmonella flagellin proteinine denk düşmektedir. Bu ön çalışmada, monoepitopik ve monoizotipik antikor salgılayan monoklonal hibrid hücrelerin elde edilmesi için ilave araştırmaların yapılmasının gerekli olduğu sonucuna varılmıştır.

Anahtar Kelimeler:

antikor, hibridoma, inaktivasyon, fareler, dalak hücreleri, Salmonella enteritidis

Anti-Salmonella Enteritidis antikorları salgılayan fare hibridomalarının geliştirilmesi ve ön karakterizasyonları

BALB/c mice were intraperitoneally immunized with inactivated bacteria for generation of monoclonal anti-S. Enteritidis antibody. The spleen cells of the highest responder animal at fifth immunization were used as the fusion partner of the mouse Sp2/0 myeloma cells. A total of 6 stable hybridomas secreting IgM and IgG isotype antibodies were obtained. These hybridomas were found to be reactive with three S. Enteritidis antigens having relative molecular weights of 73, 59 and 42kDa in Western blot analysis. The 59kDa molecule corresponds to the flagellin protein. From this preliminary study, it can be concluded that further investigations are necessary to obtain monoclonal hybrid cells secreting monoepitopic and monoisotypic antibody by subcloning of the parental hybridomas.

Keywords:

antibodies, hybridomas, inactivation, mice, spleen cells, Salmonella enteritidis,

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