title: Denatured G-protein coupled receptors as immunogens to generate highly specific antibodies.
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.
authors.affiliation: CNRS, IPBS (Institut de Pharmacologie et de Biologie Structurale), Toulouse, France. franck.talmont@ipbs.fr
authors.forename: Franck
authors.initials: F
authors.lastname: Talmont
authors.forename: Lionel
authors.initials: L
authors.lastname: Moulédous
authors.forename: Jérôme
authors.initials: J
authors.lastname: Boué
authors.forename: Catherine
authors.initials: C
authors.lastname: Mollereau
authors.forename: Gilles
authors.initials: G
authors.lastname: Dietrich
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics; D011489:Protein Denaturation; D017510:Protein Folding; D018013:Receptors, Neuropeptide / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D017473:Receptors, Opioid, kappa / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D017450:Receptors, Opioid, mu / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
chemical_list: D000906:Antibodies; D007074:Immunoglobulin G; D018013:Receptors, Neuropeptide; D017473:Receptors, Opioid, kappa; D017450:Receptors, Opioid, mu; D011994:Recombinant Proteins