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Anti-phosphorylation Tau protein antibody use for detecting AD sign abnormal phosphation Tau protein level

A phosphorylated and non-phosphorylated technology, applied in the fields of immunobiology and medical diagnosis, which can solve the problems of few chemical detection and related diagnostic methods and staying in the research scope.

Active Publication Date: 2012-04-04
南京川博生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, many companies and research institutions in the world are actively carrying out research in this area, but currently there are relatively few immunohistochemical detection and related diagnostic methods based on phosphorylation site-specific Tau protein antibodies, and most of them stay in the research category

Method used

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  • Anti-phosphorylation Tau protein antibody use for detecting AD sign abnormal phosphation Tau protein level
  • Anti-phosphorylation Tau protein antibody use for detecting AD sign abnormal phosphation Tau protein level
  • Anti-phosphorylation Tau protein antibody use for detecting AD sign abnormal phosphation Tau protein level

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1. Preparation of a class of antibodies against phosphorylated Tau protein:

[0058] (1). Antigen peptide synthesis and carrier coupling: According to the sequence information of the human Tau protein in the protein database, six key phosphorylated amino acid positions of Thr181, Thr205, Thr231, Ser262, Ser396 and Ser404 were designed respectively. The 14AA polypeptides that are continuous on both sides of the dot, including phosphate amino acids (a cysteine ​​is artificially added to the N-terminus), the specific sequences of these six antigenic polypeptides are (Note: The italicized and double-underlined sequences in the sequence are phosphorylated Amino acid):

[0059] Thr181 CTPPAPK PPSSGE

[0060] Thr205 CSPGSPG PGSRSR

[0061] Thr231 CKVAVVR PPKSPS

[0062] Ser262 CVKSKIG TENLKH

[0063] Ser396 CAEIVYK PVVSGD

[0064] Ser404 CVVSGDT PRHLSN

[0065] (2). The above six antigenic polypeptides (P peptides) containing phosphorylated amino acids at corresponding posi...

Embodiment 2

[0072] Example 2. Anti-phospho-Tau protein antibody was used to detect phosphorylated Tau protein in brain tissue of AD rat animal model by immunohistochemistry:

[0073] 1. Aβ25-35 injection to establish AD rat model:

[0074] Dissolve 1 mg of Aβ25-35 in 200μL of sterile physiological saline to make a concentration of 5nmol / μL, seal and place it in a 37°C cell incubator for 96 hours to form Aβ25-35 condensed state. Divided into three groups of drug delivery models:

[0075] AD model rats: After intraperitoneal injection of 4% pentobarbital sodium anesthetized, the rats were fixed in a flat skull position with a stereotaxic instrument, and the right bregma was determined to be 3.8 mm behind the bregma and 2.5 mm beside the midline, below the surface of the skull 3.0mm is the dorsal part of the right hippocampus. A 1μL microsyringe is used to slowly inject 25-35μL of Aβ within 5 minutes, and the needle is retained for 10 minutes. After operation, 50,000 units of penicillin sodium we...

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Abstract

The invention provides an anti-phosphorylation Tau protein antibody and an application of the antibody in Alzheimer disease abnormal phosphorylation Tau protein level determination. The antibody comprises rabbit polyclonal antibodies which are respectively anti phosphorylation Tau protein at Thr181 site, anti phosphorylation Tau protein at Thr205 site, anti phosphorylation Tau protein at Thr231site, anti phosphorylation Tau protein at Ser262 site, anti phosphorylation Tau protein at Ser396 site and anti phosphorylation Tau protein at Ser404 site. The Tau protein comprises Tau protein from human beings, Tau protein from a rat and Tau protein from a mouse. The application of the antibody in the Alzheimer disease abnormal phosphorylation Tau protein level determination comprises the following steps of: preparing a brain tissue paraffin section for each group through establishing an Alzheimer disease rat model; adopting the antibodies which respectively resist the phosphorylation Tau protein of each site as a first antibody; and detecting the phosphorylation Tau protein in nerve cells of the rat brain tissue of each group so as to detect the phosphorylation Tau protein of each site. The six antibodies can be developed into kits for diagnoses and evaluations of the Alzheimer disease for human and related animal models.

Description

Technical field [0001] The invention belongs to the category of immunobiology and medical diagnosis, and specifically relates to an antibody against the amino acid phosphorylation of Tau protein at Thr181, Thr205, Thr231, Ser262, Ser396, and Ser404, and its use in AD (Alzheimer's disease) Use of abnormal phosphorylated Tau protein level measurement. Background technique [0002] Tau protein is a phosphorous glycoprotein that is widely present in neurons, and is mainly concentrated in the axons of nerve cells in the brain (the binding force to axons is stronger than that to cell bodies or dendrites). By combining with the microtubules in the nerve axons, it can promote the polymerization of microtubules, prevent depolymerization and maintain the function of microtubules. There are at least 5-6 isoforms of tau protein isolated from the brain of healthy adults, all of which are encoded by the same single gene located on the long arm of human chromosome 17. The gene is about 100kb ...

Claims

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Application Information

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IPC IPC(8): C07K16/18C07K1/14G01N33/68
Inventor 林世康夏树开孙钵清胡云龙黄文华李保仝
Owner 南京川博生物技术有限公司