I need to compare the results from western blotting and ELISA (human samples infected and uninfected with Schistosoma mansoni). How can I do? I would like references about this. Thanks
Elisa is more straight forward, you are looking for anti-parasite antibodies in the serum of the cases. Are you making a study of the variety of antibodies or proteins released during the infection? What is the purpose of the work? That would clarify the best procedure
I agree with Clive, I think that you must be clear about what you do want to test: Antibody response or antigen released by the parasite. Indirect ELISAs are extensively used for detecting antibodies, but their specificity depends of what kind of antigens are you using. It is well known that parasite antigens show a lot of cross reactivity, for example Schistosoma mansoni and Fasciola hepatica. In this regard the Western blot can provide more information than ELISA because the antigens are separate previously by an electrophoretic step. On the other hand, if you want to detect antigens, you need having specific antibodies against to your targets. MAbs are the best option.
ELISA results are in OD values (numbers). To compare results, probably you need to quantify your western results also. To quantify them you can compare intensity of bands (densitometry, image analysis tools) which can be expressed in numbers.
Correlation may exist depending on the proteins produced (xx) and detection of same with Anti-xx Ab in ELISA. It depends on what's the exact answer you are looking for.
The conditions of your coated in ELISA and those loaded in the well in gel for Western blotting may be different. It also depends whether you are doing it in native or denatured western and the reagents used in ELISA, e.g. BSA vs. Superblock as diluent. Geberally, ELISA, I think, would allow you to calculate EC50 if true binding at certain titration of the primary antibody, but it is not very specific as in Western unless you are loading a lysate of sort. ELISA of course is more high throughput and less pain especially compared to native blue Western. You may also consider Luminex and SPR. Hope it helps.
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