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Phospho-specific antibody that recognizes tau

An antibody and specific technology, applied in the direction of antibodies, antibody medical components, specific peptides, etc., can solve unmet problems

Active Publication Date: 2021-06-25
AC IMMUNE SA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0024] For acting to eliminate amyloid pathology in AD known or presumed to cause neurodegenerative disorders, such as through, for example, intraneuronal aggregates of the hyperphosphorylated protein Tau that contribute to AD There remains an unmet need for passive and / or active immunotherapy of pathological protein conformations that are as typical as amyloid

Method used

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  • Phospho-specific antibody that recognizes tau
  • Phospho-specific antibody that recognizes tau
  • Phospho-specific antibody that recognizes tau

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0507] Example 1: Generation and screening of hybridomas and antibodies

[0508] The purpose of this study was to generate and screen anti-Tau mAbs (monoclonal antibodies). Hybridomas were generated by fusion of the spleen of a Tau vaccine immunized mouse with a myeloma cell line. The reactivity of hybridomas to phosphorylated and unphosphorylated full-length Tau proteins and phosphorylated and unphosphorylated Tau antigenic peptides used in vaccine preparation was evaluated. Hybridoma screening was also performed on Tau transgenic mouse brain sections by the reactivity of hybridoma supernatants to Tau tangles using immunochemistry.

[0509] 1.1 Method

[0510] 1.1.1 Fusion

[0511] Wild-type C57BL / 6 mice inoculated with ACI-35 (Tau 393-408 [pS396, pS404]) were used for hybridoma production. Mice were boosted again with ACI-35 vaccine on day 0 and on day 4, and fusions were performed on day 7.

[0512] 6 × 10 from immunized mice 7 (ACI-35) splenocytes with 2 × 10 7 ...

Embodiment 2

[0531] Example 2: Cloning of antibody light and heavy chain variable regions

[0532] Antibody heavy and light variable region genes from hybridoma cells were cloned and DNA sequences and positions of complementarity determining regions (CDRs) and antibody binding characteristics were determined.

[0533] Using Qiagen RNeasy Mini Kit (Cat No: 74104) from 3 x 10 6Total RNA was prepared from each hybridoma cell (1 vial). RNA was eluted in 50uL of water and examined on a 1.2% agarose gel.

[0534] V preparation with IgG and kappa constant region primers using reverse transcriptase H and V K cDNA. The first strand cDNA was amplified by PCR using a larger set of signal sequence primers. The amplified DNA was gel purified and cloned into the vector T Easy (Promega). the obtained V H and V K Clones were screened for inserts of expected size. The DNA sequences of selected clones were determined bidirectionally by automated DNA sequencing. The positions of the complementa...

Embodiment 3

[0535] Example 3: Binding Study I

[0536] The objective was to measure phospho-Tau (pTau) binding of antibodies produced from subcloned hybridomas derived from mice immunized with Tau liposomal vaccine. To perform this test, an enzyme-linked immunosorbent assay (ELISA) was used to measure the binding of purified antibodies to phosphorylated and non-phosphorylated full-length Tau protein and to phosphorylated and non-phosphorylated Tau antigenic peptides for liposomal vaccine preparation combine.

[0537] Screening is accomplished by two other methods. Immunohistochemistry (IHC) was performed on brain sections from Tau transgenic animals (TAUPIR) using an anti-Tau antibody as the primary antibody. Additionally, brain protein homogenates from Tau transgenic mice were immunoblotted (WB) using anti-Tau antibody as the blotting antibody.

[0538] 3. 1 method

[0539] 3.1.1 ELISA: Phospho-Tau binding assay

[0540] To test the binding of purified antibodies to Tau and pTa...

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Abstract

The present application relates to phosphorylation-specific antibodies that recognize Tau. The present invention relates to methods and compositions for therapeutic and diagnostic use in the treatment of diseases and conditions caused by or associated with neurofibrillary tangles, in particular, the present invention relates to the specific recognition and binding to phospho Antibodies to the Tau conformation of the pathological protein Tau, and methods and compositions related to the therapeutic and diagnostic applications of the antibodies in the treatment of Tau diseases including Alzheimer's disease (AD).

Description

[0001] This application is a divisional application of an invention application with an application date of October 5, 2012, a Chinese application number of 201280060349.3, and an invention title of "phosphorylation-specific antibody that recognizes Tau". [0002] Related applications [0003] This application claims the benefit of European Patent Application No. EP12163319.2, filed on April 5, 2012, and International Patent Application No. PCT / EP2011 / 067604, filed on October 7, 2011, the contents of each of which are hereby incorporated by reference Reference is incorporated in its entirety. [0004] Field of Invention [0005] The present invention relates to methods and compositions for therapeutic and diagnostic use in the treatment of diseases and disorders caused by or associated with neurofibrillary tangles. In particular, the present invention relates to antibodies that specifically recognize and bind to conformers of the phosphorylated pathological protein tau, and to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/18C12N15/13C12P21/02A61K39/395A61P25/28G01N33/68
CPCG01N33/6896A61K2039/505C07K2317/30C07K2317/34C07K2317/76G01N2800/2821A61K39/0005C07K16/18A61P21/00A61P25/00A61P25/16A61P25/28A61K39/3955C07K2317/33C07K2317/565C07K2317/92G01N2333/47G01N2440/14G01N2800/52
Inventor A·法伊弗A·穆斯M·皮尔格伦O·阿道夫森F·范勒芬
Owner AC IMMUNE SA
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