Is there a simple method to physically characterize the crude Ag extracted from a parasite to check whether they are proteins, polysaccharides or phospholipids like compounds? I am using crude Ag as the coating Ag for an ELISA. So, the requirement is to physically characterize the crude Ag and check the immunogenic activity of each fraction.
Dear Dr.
Some more information about your experiment would be helpful: From what Parasite do you want to obtained Crude Antigen? The physical characterization will also depend on what type of parasite and What kind of crude antigen . Generally
PDF attached as example
Good luck
Thanks. I am working on Leishmania parasite. Is it possible to have non-protein antigens of that parasite which are immunodominant? How to identify them? Because the positivity of my ELISA may be not only from the immumodominant protein-antigens.
You should separate the three groups of molecules and test the sample sequentially. First I would remove lipids from the sample using, e.g., Lipid Removal Agent from Sigma-Aldrich (http://www.sigmaaldrich.com/technical-documents/articles/analytix/lra-lipid-removal.html). After testing lipid-free fraction in your ELISA, I would go on with removal of saccharide compounds from the sample using a chemical method followed by dialysis, e.g. a protocol according to this paper: http://www.ncbi.nlm.nih.gov/pubmed/2825601 - then you should have 1) complete sample, 2) lipid-free sample, 3) lipid- and saccharide-free sample. You may also wish to remove nucleic acids from the samples - for this I would use enzymatic tools (some RNase and DNase).
Good luck!
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