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Proteomic methods promising for antigen identification.(Brief Article)
2002 JUL 17 - (NewsRx.com & NewsRx.net) -- by Maria G. Essig, MS, ELS, senior medical writer - Researchers in Austria employed proteomic analysis, the rapid systematic separation, identification, and characterization of proteins in a biological sample, to select candidate antigens for Staphylococcus aureus vaccines.
O. Vytvytska and colleagues at InterCell, Vienna, approached antigen cataloging from three angles. They collected serum samples from approximately 100 patients who were either healthy or infected with S. aureus and tested the sera for antistaphylococcal antibodies and antigenic surface proteins. Screening procedures resulted in the selection of the five healthy individuals and five infected patients whose sera had the highest antiprotein IgG responses.
The researchers also cultured the S. aureus COL strain under different conditions to identify the antigens expressed, which were followed by Western blot analysis.
Third, the group concentrated the surface proteins by digesting the S. aureus cell walls and separating them from the protoplasts.
Two-dimensional electrophoresis of 1:10,000 to 1:100,000 dilutions of the serum samples demonstrated the presence of antigenic staphylococcal proteins. Preparative 2D electrophoresis allowed the researchers to purify 21 of the proteins, which were identified through matrix-assisted laser desorption/ionization mass spectrometry and tandem mass spectrometry sequencing of tryptic ...
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