Monoclonal antibody against human neurofilament medium molecular weight subunit and preparation method and application thereof

A monoclonal antibody, molecular weight technology, used in biochemical equipment and methods, anti-animal/human immunoglobulin, analytical materials, etc., can solve the problems of high sensitivity, lack of sensitivity, inconvenient inspection and diagnosis, etc., to achieve high sensitivity , a strong specific effect

Inactive Publication Date: 2012-10-03
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These techniques not only require highly sensitive and specific antibodies, but also human tissue samples, so it is inconvenient to check and diagnose
At present, there is no NF-M detection kit in the world, and there is no anti-NF-M antibody in China, and there is a lack of anti-NF-M McAb with high sensitivity and specificity.

Method used

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  • Monoclonal antibody against human neurofilament medium molecular weight subunit and preparation method and application thereof
  • Monoclonal antibody against human neurofilament medium molecular weight subunit and preparation method and application thereof
  • Monoclonal antibody against human neurofilament medium molecular weight subunit and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Preparation of anti-NF-M monoclonal antibody

[0019] 1. Selection, synthesis and coupling of human NF-M peptide sequence

[0020] The amino acid sequence of NF-M is highly homologous in humans, rats, and mice. How to choose the antigen is crucial to whether BALB / c mice can stimulate an immune response. The antigenic determinants of proteins mostly exist at the C-terminus or N-terminus. After comparing the differences in the amino acid sequence of human, rat, and mouse NF-M, we selected an amino acid sequence at the N-terminus of the NF-M amino acid sequence. GNPSAYRRVTETRSSFSRVSGSPSSGFRSQSWSRGSP-C, synthetic peptide, Sulfo-SMCC method coupled with KLH to prepare complete antigen NF-M-KLH, the coupling rate was 92.37%.

[0021] 2. Animal immunity

[0022] The NF-M-KLH conjugate has high coupling efficiency, stable coupling antigen, good solubility, and strong immunogenicity.

[0023] The NF-M-KLH conjugate was formulated into a 1mg / ml solution, and a small-dose long-p...

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Abstract

The invention discloses a monoclonal antibody against a human neurofilament medium molecular weight subunit and a preparation method and application thereof. The monoclonal antibody has specificity for polypeptide with an amino acid sequence shown in SEQ ID NO.1 and is produced by secretion of a cell strain with the preservation number CGMCC No. 6064. The invention also provides a hybridoma cell strain for production of the monoclonal antibody, which is named as 2C1 and has been preserved in China General Microbiological Culture Collection Center on April 27, 2012, and the preservation number is CGMCC No. 6064. The monoclonal antibody has high sensitivity, high specificity and cross-reactivity with neurofilaments of mouse and rats, and can be applied in research of detection kits for neurofilament medium molecular weight subunits of human beings, mouse and rats.

Description

Technical field [0001] The invention relates to a monoclonal antibody against the molecular weight subunit of human neurofilament, a preparation method and application thereof, and a hybridoma cell strain secreting and producing the monoclonal antibody, belonging to the technical field of bioengineering. Background technique [0002] Neurofilament (Neurofilament, NF) is a specialized intermediate filament that exists in the cell body and axon of nerve cells. It has a fibrous structure with an outer diameter of about 10 nm. It is composed of high molecular weight (NF-H) and medium molecular weight (NF-H). M) and low molecular weight (NF-L) are composed of three subunits, whose molecular weights in mammals are about 200KD, 160KD and 68KD, respectively. The physiological function of NF is related to the material transportation in nerve cells and the maintenance of axon shape, diameter and extension. [0003] NF is related to neurological diseases caused by many reasons. Lateral scle...

Claims

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Application Information

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IPC IPC(8): C07K16/18C12N5/20G01N33/68G01N33/577
Inventor 赵丽谢克勤董华邵晓颖杨曦伟
Owner SHANDONG UNIV
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