Use of monoclonal antibodies to distinguish protein...

C - Chemistry – Metallurgy – 07 – K

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C07K 16/28 (2006.01) A61K 39/395 (2006.01) C12N 5/12 (2006.01) C12Q 1/02 (2006.01) G01N 33/53 (2006.01) G01N 33/566 (2006.01)

Patent

CA 2502281

Methods of preparing monoclonal antibodies that differentially bind to a single conformer of a protein of interest are described. Passive immunization using these antibodies as well as use of conformer-specific antibodies as diagnostic reagents for the purpose of stratification of patient populations with regards to disease outcome, drug efficacy or drug sensitivity is also disclosed as well as active immunization with the protein conformer. In the screening techniques, detection can be for example by tissue immunostaining, western blotting or solution IP. A specific mab termed 7VC which shows conformation specificity to CtmPrP, a prion protein conformer that triggers neurodegeneration under specific assay conditions of pH and copper concentration, is described. A second specific antibody termed 19B10 shows conformation specificity for NtmPrP, a prion protein conformer that downregulates total PrP expression and effects cell differentiation.

L'invention concerne des méthodes permettant de préparer des anticorps monoclonaux qui se lient de manière différentielle à un seul conformère d'une protéine d'intérêt. L'invention concerne également l'immunisation passive à l'aide de ces anticorps; l'utilisation d'anticorps spécifiques d'un conformère comme réactifs diagnostic à des fins de stratification de populations de patients afin de mesurer le résultat d'une maladie, l'efficacité d'un médicament ou sa sensibilité; et l'immunisation active à l'aide du conformère de protéine. Dans les techniques de criblage, la détection peut s'effectuer, par exemple, par immunocoloration d'un tissu, par transfert western ou à l'aide d'une solution IP. L'invention concerne, en outre, un anticorps monoclonal spécifique appelé 7VC qui montre une spécificité de conformation avec CtmPrP, un conformère de protéine prion qui déclenche une neurodégénération dans des conditions de dosage spécifiques de pH et de concentration de cuivre. L'invention concerne enfin un second anticorps spécifique appelé 19B10 montrant une spécificité de conformation avec NtmPrP, et un conformère de protéine prion réduisant l'expression totale de PrP et effectuant une différentiation cellulaire.

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