Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis.
Autor(es): Fontaine A; Pascual A; Diouf I; Bakkali N; Bourdon S; Fusai T; Rogier C; Almeras L
Resumo: Mosquito salivary proteins are involved in several biological processes that facilitate their blood feeding and have also been reported to elicit an IgG response in vertebrates. A growing number of studies have focused on this immunological response for its potential use as a biological marker of exposure to arthropod bites. As mosquito saliva collection is extremely laborious and inefficient, most research groups prefer to work on mosquito salivary glands (SGs). Thus, SG protein integrity is a critical factor in obtaining meaningful data from immunological and biochemical analysis. Current methodologies rely on an immediate freezing of SGs after their collection. However, the maintenance of samples in a frozen environment can be hard to achieve in field conditions. In this study, SG proteins from two mosquito species (Aedes aegypti and Anopheles gambiae s.s.) stored in different media for 5 days at either +4°C or room temperature (RT) were evaluated at the quantitative (i.e., ELISA) and qualitative (i.e., SDS-PAGE and immunoblotting) levels. Our results indicated that PBS medium supplemented with an anti-protease cocktail seems to be the best buffer to preserve SG antigens for 5 days at +4°C for ELISA analysis. Conversely, cell-lysis buffer (Urea-Thiourea-CHAPS-Tris) was best at preventing protein degradation both at +4°C and RT for further qualitative analysis. These convenient storage methods provide an alternative to freezing and are expected to be applicable to other biological samples collected in the field.
Palavras-Chave: Salivary Gland; Mosquito Species; Cell Lysis Buffer; Salivary Protein; Preservation Condition
Imprenta: Parasites & Vectors, v. 8, n. 4:33, 2011
Identificador do objeto digital: 10.1186/1756-3305-4-33.
Descritores: Aedes aegypti - Cell ; Aedes aegypti - Pathogenesis ; Aedes aegypti - Proteins ; Aedes aegypti - Immunology
Data de publicação: 2011