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Human occludin antigen epitope peptide, antigen, antibody, kit and use

A protein antigen, epitope peptide technology, applied to the antibody in the in vitro diagnostic kit for human atresia, sex antigen and corresponding monoclonal antibody or polyclonal antibody, the field of human atresia, can solve the BBB Barrier function damage and other problems, to achieve the effect of assessing the degree of damage and reducing serious consequences

Active Publication Date: 2018-05-29
深圳市安群生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, once cerebral ischemia and reperfusion after ischemia occur, the barrier function of the BBB will be gradually destroyed

Method used

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  • Human occludin antigen epitope peptide, antigen, antibody, kit and use
  • Human occludin antigen epitope peptide, antigen, antibody, kit and use
  • Human occludin antigen epitope peptide, antigen, antibody, kit and use

Examples

Experimental program
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preparation example Construction

[0037] The preparation method of the occludin antigenic epitope peptide of the present invention can be a chemical synthesis method: the antigenic epitope peptide is synthesized by a solid-phase method using an American ABI431A type polypeptide automatic synthesizer. The molecular weights of the antigenic epitope peptides (1) and (2) of the present invention are 1971.13 and 2124.44 respectively, which can be determined by mass spectrometry, and the synthesized antigenic epitope peptide sequences are identified by polypeptide sequence determination. The purity of the peptides was evaluated by thin-layer chromatography and high-performance liquid chromatography, and the concentration of the epitope peptide was determined.

[0038] 2. Occlusin antigen

[0039] The present invention also provides an atretin antigen prepared by coupling one of the human atretin epitope peptides (1) and (2) of the present invention with a carrier protein. Specifically, the present invention provide...

Embodiment 1

[0054] Example 1: Preparation of Octrein epitope peptides (1) and (2).

[0055] The preparation method uses chemical synthesis method: using the American ABI431A automatic peptide synthesizer, the occludin epitope peptides (1) and (2) are synthesized respectively by solid-phase method. The purity of the epitope peptide was assessed by high performance liquid chromatography, and the concentration of the peptide was determined. The molecular weights of the antigenic epitope peptides (1) and (2) of the present invention are 1971.13 and 2124.44 respectively, which are determined by mass spectrometry, and the synthesized polypeptide sequences are identified by polypeptide sequence determination.

[0056] 1. Synthesis of Occlusin Epitope Peptides (1) and (2)

[0057] The above peptides were synthesized by solid-phase method. The main idea of ​​solid-phase peptide synthesis is: first connect the carboxyl group of the carboxyl-terminal amino acid of the peptide chain to be synthesiz...

Embodiment 2

[0145] Example 2: The atretin antigen epitope peptides (1) and (2) obtained in Example 1 were linked to carrier proteins to prepare atretin antigens (1) and (2), and the obtained antigens (1) and (2) were used ) to immunize animals respectively, thereby using the antigen (1) to prepare specific monoclonal antibodies and polyclonal antibodies, and using the antigen (2) to prepare specific monoclonal antibodies and polyclonal antibodies.

[0146] 1. Antigen preparation: Occlusin peptides (1) and (2) were respectively linked with carrier protein KLH (keyhole limpet hemocyanin) (obtained from sigma company) by BDB (Bis-diazotizedbenzidine dichloride) method to prepare atocrin Antigens (1) and (2).

[0147] Take 10.0mg of Occlucin peptide (1) or (2), dissolve it with 1ml 0.1M PBS buffer (pH 7.4); dissolve 10mg of KLH with 20ml of 0.2M borate buffer (pH 9.0); then mix the two , cooled to 0°C, take BDBCl 2 110 μL, reacted at room temperature for 1.5 h, dialyzed overnight, then ali...

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Abstract

The present invention relates to a human occludin antigen epitope peptide, a human occludin antigen, a human occludin antibody, a kit and use. An amino acid sequence of the human occludin antigen epitope peptide is as shown in one of sequence table SEQ ID NO. 1 and sequence table SEQ ID NO.2. The human occludin antigen is prepared from the human occludin antigen epitope peptide and a protein carrier by coupling. A human occludin monoclonal or polyclonal antibody is prepared from the human occludin antigen. The human occludin monoclonal or polyclonal antibody is used for the preparation of an occludin in-vitro diagnostic kit. The human occludin antigen epitope peptide has good antigenicity, highly specific monoclonal and polyclonal antibodies can be produced by use of an antigen (immunogen)produced from the human occludin antigen epitope peptide for immunizing an animal, and the human occludin antigen epitope peptide can be applied to human occludin in vitro testing.

Description

technical field [0001] The invention belongs to the field of polypeptide chemistry and immunology, and in particular relates to a human occludin epitope peptide, an occludin-specific antigen prepared by using the epitope peptide, a corresponding monoclonal antibody or a polyclonal antibody, and the antibody The application in the preparation of the human atretin in vitro diagnostic kit, the human atretin in vitro diagnostic kit, and the application of the kit in the quantitative detection of human atretin in serum. Background technique [0002] The blood brain barrier (BBB) ​​refers to the barrier between plasma and brain cells formed by brain capillary walls and glial cells, and the barrier between plasma and cerebrospinal fluid formed by the choroid plexus. Some substances (mostly harmful) enter the brain tissue from the blood. The BBB is composed of capillary endothelial cells, astrocytes, some neurons, peritubular cells, tight junctions between vascular endothelial cell...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/705C07K16/28G01N33/68G01N33/577
CPCC07K14/705C07K16/28G01N33/577G01N33/68G01N2333/705G01N2800/2871
Inventor 朱建安朱仕杰
Owner 深圳市安群生物工程有限公司
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