ABSTRACT
Objective To identify antigenic proteins from the salivary glands of female Anopheles maculatus using a proteomic approach to find the biomarker candidate for serological tools. Methods The identification of antigenic proteins of Anopheles maculatus salivary gland used these techniques: one-dimensional gel electrophoresis (sodium dodecyl sulfate polyacrylamide gel electrophoresis), western blot, and liquid chromatography–mass spectrometry. Results The proteins that have molecular weight (MW) 43 and 34 kDa were the antigenic protein. Computational bioinformatic analysis by Mascot Server revealed seven novel hypothetical proteins (MW: 43 kDa) and two novel hypothetical proteins (MW: 34 kDa). Further analysis (BLASTP, antigenicity, epitope mapping, and specificity analysis) showed that two novel proteins were identified as apolipoprotein D and cathepsin D in Anopheles darlingi. Conclusions The identified proteins are potential to be developed as a biomarker of mosquito bite's exposure.
ABSTRACT
Humans develop anti-salivary proteins after arthropod bites or exposure to insect salivary proteins. This reaction indicates that vector bites have a positive effect on the host immune response, which can be used as epidemiological markers of exposure to the vector. Our previous study identified two immunogenic proteins with molecular weights of 31 kDa and 56 kDa from salivary gland extract [SGE] of Aedes aegypti that cross-reacted with serum samples from Dengue Hemorrhagic Fever [DHF] patients and healthy people in an endemic area [Indonesia]. Serum samples from individuals living in non-endemic area [sub-tropical country] and infants did not show the immunogenic reactions. The objective of this research was to identify two immunogenic proteins, i.e., 31 and 56 kDa by using proteomic analysis. In this study, proteomic analysis resulted in identification of 13 proteins and 7 proteins from the 31 kDa- and 56 kDa-immunogenic protein bands, respectively. Among those proteins, the D7 protein [Arthropode Odorant-Binding Protein, AOBP] was the most abundant in 31-kDa band, and apyrase was the major protein of the 56-kDa band