Electroelution of 31 kDa Immunogenic Protein Fraction from the Salivary Gland of Aedes aegypti and Aedes albopictus (Diptera: Culicidae)

Abstract

Dengue Hemorrhagic Fever (DHF) is a disease generated by dengue virus infection. The dengue virus is transmitted to the host by mosquito vectors. The main and secondary vectors are Aedes aegypti and Aedes albopictus. Dengue virus transmission is mediated by salivary gland proteins that facilitate the blood feeding process. Previous studies have shown that the molecular weight of 31 kDa is an immunogenic protein of Ae. aegypti belonging to the D7 family based on the result of Mass Spectrometry. This immunogenic protein can be used for the development of vector-based dengue vaccines. The purpose of this study is to purify the 31 kDa protein fraction from the salivary gland of Ae. aegypti and Ae. albopictus using the electroelution method. The study is conducted by collecting the salivary glands of Ae. aegypti and Ae. albopictus, SDS-PAGE, electroelution using the Bio-Rad 422 electroeluter, Dot Blot, and Western Blot. Both protein samples resulting from electroelution confirmed with a single band appeared in SDS-PAGE. The optimal conditions for electroelution are 3 h running time, 100 voltage, volume ±1 mL, and the concentration obtained are 1.789 mg/mL (Ae. aegypti) and 1.81 mg/mL (Ae. albopictus). These results are supported by the other result of the dark dots shown in Dot Blot and a single band of 31 kDa shown in Western Blot when it reacted with the serum of dengue patients, endemic healthy people, and neonates. These results indicate that the purified 31 kDa immunogenic protein fraction can be recognized by specific antibodies.