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Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis
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 ex...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3068118/ https://www.ncbi.nlm.nih.gov/pubmed/21385450 http://dx.doi.org/10.1186/1756-3305-4-33 |
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author | Fontaine, A Pascual, A Diouf, I Bakkali, N Bourdon, S Fusai, T Rogier, C Almeras, L |
author_facet | Fontaine, A Pascual, A Diouf, I Bakkali, N Bourdon, S Fusai, T Rogier, C Almeras, L |
author_sort | Fontaine, A |
collection | PubMed |
description | 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. |
format | Text |
id | pubmed-3068118 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-30681182011-03-31 Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis Fontaine, A Pascual, A Diouf, I Bakkali, N Bourdon, S Fusai, T Rogier, C Almeras, L Parasit Vectors Short Report 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. BioMed Central 2011-03-08 /pmc/articles/PMC3068118/ /pubmed/21385450 http://dx.doi.org/10.1186/1756-3305-4-33 Text en Copyright ©2011 Fontaine et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Short Report Fontaine, A Pascual, A Diouf, I Bakkali, N Bourdon, S Fusai, T Rogier, C Almeras, L Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis |
title | Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis |
title_full | Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis |
title_fullStr | Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis |
title_full_unstemmed | Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis |
title_short | Mosquito salivary gland protein preservation in the field for immunological and biochemical analysis |
title_sort | mosquito salivary gland protein preservation in the field for immunological and biochemical analysis |
topic | Short Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3068118/ https://www.ncbi.nlm.nih.gov/pubmed/21385450 http://dx.doi.org/10.1186/1756-3305-4-33 |
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