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PLoS One. 2012;7(9):e46348. doi: 10.1371/journal.pone.0046348. Epub 2012 Sep 27.

Denatured G-protein coupled receptors as immunogens to generate highly specific antibodies.

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1
CNRS, IPBS (Institut de Pharmacologie et de Biologie Structurale), Toulouse, France. franck.talmont@ipbs.fr

Abstract

G-protein coupled receptors (GPCRs) play a major role in a number of physiological and pathological processes. Thus, GPCRs have become the most frequent targets for development of new therapeutic drugs. In this context, the availability of highly specific antibodies may be decisive to obtain reliable findings on localization, function and medical relevance of GPCRs. However, the rapid and easy generation of highly selective anti-GPCR antibodies is still a challenge. Herein, we report that highly specific antibodies suitable for detection of GPCRs in native and unfolded forms can be elicited by immunizing animals against purified full length denatured recombinant GPCRs. Contrasting with the currently admitted postulate, our study shows that an active and well-folded GPCR is not required for the production of specific anti-GPCR antibodies. This new immunizing strategy validated with three different human GPCR (μ-opioid, κ-opioid, neuropeptide FF2 receptors) might be generalized to other members of the GPCR family.

PMID:
23029489
PMCID:
PMC3459905
DOI:
10.1371/journal.pone.0046348
[Indexed for MEDLINE]
Free PMC Article
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