Proteomic analysis of africanized bee venom: a comparison of protein extraction methods

Abstract

The Africanised bee is the most common type of bee in Colombia, and therapeutic properties for different diseases have been attributed to its venom, without much scientific support. A literature search of reports on the proteomic analysis of honeybee venom yielded four different methods for extracting proteins from bee venom. The first method consists in resuspending the venom in 7 M Urea, followed by precipitation with acetone and finally resuspending the pellet in 7 M Urea and 4 % CHAPS. For the second method, the venom is resuspended in lysis buffer, precipitated with trichloroacetic acid, and then resuspended in 7 M Urea and 4 % CHAPS. The third method is similar to the previous one, except that the precipitation step is performed with acetone instead of trichloroacetic acid. Finally, the fourth method is to resuspend the venom in distilled water, precipitate with acetone and resuspend in 7 M Urea and 4 % CHAPS. This work focused on comparing the performance of these four extraction methods, in order to determine the method with the best results in terms of concentration and integrity of the proteins obtained. Of the four methods evaluated, the best results in terms of protein concentration and yield were obtained by resuspending the bee venom in lysis buffer followed by precipitation with acetone (method 3), and by resuspending in distilled water followed by precipitation with acetone (method 4). Of these, the method that maintained protein integrity and yielded the best proteomic profile was that in which the bee venom was resuspended in lysis buffer followed by precipitation with acetone (method 3).

Keywords

• acetona
• apitoxina
• electroforesis en gel bidimensional
• perfil proteíco.