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Hybridoma cell strain secreting anti-serine phosphorylated tau protein monoclonal antibody

A hybridoma cell line, serine phosphorylation technology, applied in the field of immunodetection, can solve the problems of less research and other issues

Active Publication Date: 2021-06-22
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, more phosphorylated tau proteins are studied, including p-Tau 181 , p-Tau 231 , p-Tau 199 , while p-Tau 396 , p-Tau 404 less research
The monoclonal antibody and its detection kit targeting serine phosphorylated tau protein at positions 396 and 404 simultaneously have not been reported yet

Method used

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  • Hybridoma cell strain secreting anti-serine phosphorylated tau protein monoclonal antibody
  • Hybridoma cell strain secreting anti-serine phosphorylated tau protein monoclonal antibody
  • Hybridoma cell strain secreting anti-serine phosphorylated tau protein monoclonal antibody

Examples

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Embodiment 1

[0019] Example 1: Anti-human p-Tau 396,404 Preparation of specific monoclonal antibodies

[0020] For the convenience of reference below, the meanings of the main abbreviations are explained as follows:

[0021] p-Tau 396,404 Monomer, i.e., p-Tau 396,404 Monomer: A single protein formed from human Tau protein phosphorylated at serine 396 and 404.

[0022] p-Tau 396,404 aggregates, i.e., p-Tau 396,404 Oligomer: Human Tau protein phosphorylated at serine 396 and 404 is obtained by mutual folding.

[0023] 1. Antigen preparation

[0024] The Tau protein (p-Tau 396,404 ), dissolved in TBS solution to a final concentration of 1 mg / mL. Result: The results of SDS-PAGE showed that the synthetic peptide was of good purity and the electrophoresis bands were uniform ( figure 1 in A).

[0025] 2. Preparation of mouse monoclonal antibody

[0026] 2.1. Immunization of mice

[0027] Mix 100 μL of the prepared antigen with an equal volume of Freund’s complete adjuvant for the firs...

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Abstract

The invention belongs to the field of immunodetection, and discloses two hybridoma cell strains secreting anti-serine phosphorylation tau protein monoclonal antibodies, and the hybridoma cell strains used for secreting p-Tau 396, 404 monoclonal antibodies, the hybridoma cell strains are preserved in the China Center for Type Culture Collection (CCTCC), and the preservation numbers are respectively CCTCC NO: C2020219 and CCTCC NO: C2020218. According to the invention, two hybridoma cell strains capable of secreting novel p-Tau 396, 404 monoclonal antibodies are provided, and the monoclonal antibodies capable of specifically recognizing human p-Tau 396, 404 and aggregates thereof can be prepared by using the hybridoma cell strains, so that the problem of lack of antibodies capable of simultaneously targeting p-Tau 396, 404 at present is solved. The two monoclonal antibodies provided by the invention can be further applied subsequently to prepare a high-specificity and high-sensitivity p-Tau 396, 404 detection kit, the immunological treatment of specific targeting p-Tau 396, 404 is developed, the disease progress is slowed down, and the blank of the current immunological treatment of targeting p-Tau 396, 404 is filled up.

Description

technical field [0001] The present invention belongs to the field of immunoassay, and more specifically, relates to two kinds of hybridoma cell lines secreting anti-serine phosphorylated tau protein monoclonal antibody, that is, two kinds of Tau protein capable of secreting anti-human phosphorylated serine at 396 and 404 positions (p-Tau 396,404 ) Hybridoma cell lines of monoclonal antibodies. Background technique [0002] Alzheimer's disease (AD) is a neurodegenerative disease with insidious onset and progressive development. Clinically, it is characterized by comprehensive dementia such as memory impairment, aphasia, apraxia, agnosia, impairment of visuospatial skills, executive dysfunction, and personality and behavior changes. The etiology of most Alzheimer's diseases is still unknown, and only Genetic differences have been identified in 1% to 5% of cases. Alzheimer's disease is the most common cause of dementia and is responsible for 60% to 80% of dementia cases worl...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/18C07K16/44G01N33/68G01N33/577A61K39/395A61P25/28
CPCC07K16/18C07K16/44G01N33/6896G01N33/577A61P25/28C07K2317/92C07K2317/76G01N2333/47G01N2800/2821
Inventor 骆海明张立定李艳青牛是琦
Owner HUAZHONG UNIV OF SCI & TECH
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