A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte

A new method for the capture of surface proteins in Plasmodium falciparum parasitized erythrocyte

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Introduction: We propose a new method for the selective labeling, isolation and electrophoretic analysis of the Plasmodium falciparum protein Collections exposed on the erythrocyte cell surface.Historically, membrane surface proteins have been isolated using a surface biotinylation followed by capture of biotin-conjugated protein via an avidin/streptavidin-coated solid support.The major drawback of the standard methods has been the labeling of internal proteins due to fast internalization of biotin.

Methodology: To solve this problem, we used a biotin label that does not permeate through the membrane.As a further precaution to avoid the purification of non surface exposed proteins, we directly challenged whole labeled cells with avidin coated beads and then solubilized them using non ionic detergents.Results: A marked enrichment of most of the RBC membrane proteins known to face the external surface of the membrane validated the specificity of the method; furthermore, only small amounts of haemoglobin and cytoskeletal proteins were detected.

A wide range of P.falciparum proteins were additionally described to be exposed on the erythrocyte surface.Some of them have been previously observed and used as vaccine candidates while a number of newly described antigens have been presently identified.

Those antigens require further characterization and validation with additional methods.Conclusion: Surface proteins preparations were very reproducible and Waterwear Salopettes identification of proteins by mass spectrometry has been demonstrated to be feasible and effective.