denatured g-protein coupled receptors as immunogens to generate highly specific antibodies变性蛋白的受体产生高度特定抗体的免疫原.pdfVIP

Denatured G-Protein Coupled Receptors as Immunogens to Generate Highly Specific Antibodies 1,2 ´ 1,2 ´ ˆ ´ 2,3,4 1,2 2,3,4 Franck Talmont *, Lionel Mouledous , Jerome Boue , Catherine Mollereau , Gilles Dietrich ´ 1 CNRS, IPBS (Institut de Pharmacologie et de Biologie Structurale), Toulouse, France, 2 Universite Paul Sabatier Toulouse III, Toulouse, France, 3 INSERM, U1043, Centre de Physiopathologie de Toulouse-Purpan, Toulouse, France, 4 CNRS, U5282, Toulouse, France 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 (m-opioid, k-opioid, neuropeptide FF2 receptors) might be generalized to other members of the GPCR family. ´ ´ Citation: Talmont F, Moul