Implementação de uma metodologia para a separação do proteoma do plasma humano por eletroforese bidimensional

The conditions to separate blood plasma proteomes by two-dimensional electrophoresis were implemented. In plasma samples from infant and adult two sample pretreatment systems to reduce the proteins dynamic range were evaluated: Immunodepletion of abundant proteins and protein-enrichment of low abundance proteins. Proteomes were separated by two-dimensional electrophoresis and the images were analyzed using Melanie 7.0. It was found that both pretreatment methods were reproducible and allowed to see the differences in the proteomes of infant and adult, as evidenced by the principal component analysis and heatmap, a type of hierarchic classification. The percent recovery of proteins was greater with the immunodepletion method, compared to the protein enrichment system. With these results, we conclude that reducing the complexity of plasma by immunoaffinity showed better control of unrecovered proteins and therefore less loss of information, which allows its application on exploratory studies to identify potential biomarkers of disease.