4aβ1‑15 derived monoclonal antibody and its application

A monoclonal antibody, antibody technology, applied in the field of immunology

Active Publication Date: 2017-06-20
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, passive immunization results from a preclinical study of aged APP23 mice receiving weekly N-terminal-directed anti-Aβ1-42 antibodies for 5 months revealed treatment-related side effects

Method used

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  • 4aβ1‑15 derived monoclonal antibody and its application
  • 4aβ1‑15 derived monoclonal antibody and its application
  • 4aβ1‑15 derived monoclonal antibody and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 14

[0051] Example 1 Synthesis of 4Aβ1-15 peptide

[0052] For the preparation method of 4Aβ1-15 peptide, please refer to the patent No. "ZL 201010163165.2" and the invention patent named "High-efficiency secretion expression and purification method of recombinant 4Aβ15 in Pichia pastoris", wherein 4Aβ15 is the 4Aβ1-15 of the present invention . The 4Aβ1-15 polypeptide antigen used in the present invention is provided by Livzon Pharmaceutical Company, and the protein purity is >95%.

[0053] In addition, the full-length 4Aβ1-15 peptide can also be artificially synthesized according to the sequence of 4Aβ1-15, wherein:

[0054] The amino acid sequence of Aβ1-15 peptide is: DAEFRHDSGYEVHHQ;

[0055] The amino acid sequence of the 4Aβ1-15 peptide is: 4 repeating sequences of DAEFRHDSGYEVHHQ, connected by the peptide segment GGPPG in the middle, with a total of 75 amino acids.

Embodiment 2

[0056] Example 2 Production of hybridoma and preparation of monoclonal antibody

[0057] animal immunity

[0058] 6-week-old Balb / c mice were immunized with purified 4Aβ1-15 antigen (protein purity>95%). The way of immunization is subcutaneous or intraperitoneal injection. For the first immunization, the antigen was emulsified with Freund's complete adjuvant (FCA; Sigma), and the immunization dose was 50 μg per mouse. After an interval of two weeks, the antigen was emulsified with Freund's incomplete adjuvant (FIA; Sigma) for the second immunization with a dose of 50 μg per mouse. Two weeks after the second immunization, the antigen was emulsified with Freund's incomplete adjuvant (FIA; Sigma) for the third immunization, and the immunization dose was 50 μg per mouse. Two weeks after the third immunization, a booster immunization was carried out, and the immunization dose was 100 μg / bird. Among them, the polypeptide antigen is dissolved in PBS at 1 mg / ml, and the adjuvant...

Embodiment 3

[0089] Example 3 Immunology, Pathology and Behavioral Observation Tests of AD Transgenic Mice Inoculated with Monoclonal Antibody

[0090] 1. Materials and methods

[0091] 1.1 Grouping and immunization procedures

[0092] The mice were divided into 4 groups (8 in each group), three of which were AD transgenic model mice APP / PS1 provided by the Model Animal Research Center of Nanjing University, and the other group were wild-type C57Bl / 6J mice without treatment. Among the three groups of transgenic mice, one group was inoculated with the monoclonal antibody 5C8H5 (hereinafter referred to as "5C8H5") as the experimental group. Since 5C8H5 was identified as an IgG phenotype, we chose mouse IgG, an irrelevant isotype control antibody, as a negative control, that is, the second group was vaccinated with mouse IgG. At the same time, we established a group of mice inoculated with 4Aβ1-15 antigen to compare whether 4Aβ1-15 and 5C8H5 under the same immunization procedure, that is,...

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Abstract

The invention discloses a monoclonal antibody derived from 4Abeta1-15. The monoclonal antibody is generated by a hybridoma cell strain with the preservation number of CCTCC C201473. The monoclonal antibody disclosed by the invention can be used for eliminating Abeta deposition without leading to micro-bleeding burden, and meanwhile, fewer microglial cells are activated so as to prevent inflammatory factors and T cells from being further excessively activated, so that the monoclonal antibody can be used for preventing and treating Alzheimer's disease.

Description

technical field [0001] The present invention relates to the field of immunology, and in particular to a monoclonal antibody derived from 4Aβ1-15, a hybridoma producing the monoclonal antibody, a pharmaceutical composition containing the monoclonal antibody, a kit containing the monoclonal antibody, and the Application of monoclonal antibodies. Background technique [0002] Alzheimer's disease (AD) is the most common form of dementia and neurodegenerative disease, causing progressive loss of memory and cognitive function in older adults. It is well known that the neuropathology of AD is characterized by plaque deposition of extracellular proteins in the brain. Both familial and sporadic cases believe that Aβ plaque is a common pathological feature, suggesting that our treatment methods should start from three aspects: inhibiting its production, promoting its degradation and enhancing its clearance from the brain. [0003] Now, stimulating the clearance of Aβ in the brain of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/18C12N5/20A61K39/395A61P25/28
Inventor 姚志彬
Owner SUN YAT SEN UNIV
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