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Help on ELISA antibody binding principles - (May/06/2005 )


May I know does ELISA bind to a native protein only or can it detect a denatured protein?

I have a self purified protein which has low antibody detection limit for ELISA.

Could it be possible if it is caused by the protein losing certain conformation during the process of purification leading to this low binding affinity?

Would really appreciate it if you could recommend me some good links or books on this topic. Thanks!



You don't need to specifics on ELISA or Western-blotting (see your other thread), as both work basically the same way, you just need information about B-cell epitopes themselves to understand the differences of antibody binding to native or denatureted protein. therefore I may recommend to you the following two papers:

1: Mahler M, Bluthner M, Pollard KM.
Advances in B-cell epitope analysis of autoantigens in connective tissue
Clin Immunol. 2003 May;107(2):65-79. Review.
PMID: 12763475 [PubMed - indexed for MEDLINE]

2: Routsias JG, Tzioufas AG, Moutsopoulos HM.
The clinical value of intracellular autoantigens B-cell epitopes in systemic
rheumatic diseases.
Clin Chim Acta. 2004 Feb;340(1-2):1-25. Review.
PMID: 14734193 [PubMed - indexed for MEDLINE]

just ignore that they are talking about autoantibodies, both discuss the different types of B-cell epitopes and methods to detect them...



Thank you very much for your help! Will get these papers ASAP. smile.gif Have a good day everyone!