Generation of monoclonal antibodies specifi c to Coenzyme A. Methods. Hybridoma technique. KLH carrier protein conjugated with CoA was used for immunization. Screening of positive clones was performed with BSA conjugated to CoA. Results. Monoclonal antibody that specifi cally recognizes CoA and CoA derivatives, but not its precursors ATP and cysteine has been generated. Conclusion. In this study, we describe for the fi rst time the production and characterization of monoclonal antibodies against CoA. The monoclonal antibody 1F10 was shown to recognize specifi cally CoA in Western blotting, ELISA and immunoprecipitation. These properties make this antiboby a particularly valuable reagent for elucidating CoA function in health and disease. K e y w o r ds: CoA, hybridoma technique, monoclonal antibody.
Aims. We aimed to study the influence of the expression level, activity and subcellular localization of CoA Synthase (CoASy) on anchoring independent growth and viability of in vitro cultured cells. Methods. Abilities of cells to form colonies in semisolid agarose and survive in growth factor depleted conditions were tested. A panel of HEK293 stable cell lines which over express wild type, catalytically inactive or mitochondria association mutants of CoASy were used in the study. Effects of CoASy down regulation by siRNA on malignant phenotype of HepG2 cells have been studied. Results. We report here that changes in CoASy expression level affect anchoring independent growth and viability of mammalian cells. Catalytic activity of CoASy and its association with mitochondria are crucial for mediating of the observed effects. Conclusions. Presented data indicate CoASy has positive impact on activity of signalling pathways in the cell and reveal unknown before functional link between signal transduction and metabolism
Відомо, що такі властивості, як наявність плазмід та синтез бактеріоциногенних речовин, у багатьох випадках мають опосередковане або пряме відношення до механізмів патогенезу, тому їхнє вивчення часто дає можливість зробити певні припущення щодо останніх та спрямувати наступні дослідження. Виявлено, що дані патогенні ізоляти з липи (Tilia sp.) мають плазміду однакової електрофоретичної рухливості, стійкі до високих концентрацій ряду антибіотиків різних типів та мають бактеріоциногенні властивості.
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