PROTEOMIC ANALYSIS OF APIS MELLIFERA VENOM DETERMINED BY LIQUID CHROMATOGRAPHY (LC) COUPLED WITH NANO-LC-MALDI-TOF/TOF MS.

The integration of multidimensional liquid chromatography and mass spectrometry analytical plat- form was proposed for proteomic exploration of honeybee venom. The combination of HPLC with nanoLC-MALDI-TOF/TOF MS system was our method of choice for compressing the dynamic range of honeybee venom protein concentration. Honeybee venom samples were separated into 6 fractions using HPLC and further analyzed by nanoLC-MALDI-TOF/TOF. Applied approach allowed to identify in total 394 peptides giving the identification of 50 components including putative toxins and trace elements. Moreover, all 12 known honeybee venom allergens were acknowledged. Additionally, four novel hypothetical proteins have been observed which were not observed in other studies. The newly recognized proteins should be further investigated, in order to characterize their functions in the venom of Apis mellifera.