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Neutralizing monoclonal antibody 12C11 of human nerve growth factor and its hybridoma cell strain

A monoclonal antibody and hybridoma cell technology, applied in the field of immunology, can solve the problems of anti-rhNGF monoclonal antibody neutralization activity identification, etc., and achieve the effect of good specificity, high titer and strong specificity

Inactive Publication Date: 2015-09-16
北京福睿君安科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that anti-rhNGF monoclonal antibody is obtained by intrasplenic injection of NGF (antigen: bacteria 1:5) adsorbed and purified by acid-treated Salmonella bacteria, and splenocytes and myeloma cells are screened after fusion with PEG. Hybridoma cells, and preliminary identification and analysis of the obtained anti-rhNGF monoclonal antibody (Su Jin, Zhang Yali, You Changxuan, et al. Preparation and preliminary identification of anti-human NGF monoclonal antibody. Journal of Cellular and Molecular Immunology. 2002,18(2):168), but no neutralizing activity of anti-rhNGF monoclonal antibody was identified

Method used

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  • Neutralizing monoclonal antibody 12C11 of human nerve growth factor and its hybridoma cell strain
  • Neutralizing monoclonal antibody 12C11 of human nerve growth factor and its hybridoma cell strain
  • Neutralizing monoclonal antibody 12C11 of human nerve growth factor and its hybridoma cell strain

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Experimental program
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Effect test

Embodiment 1

[0047] Example 1. Preparation of anti-hNGF monoclonal antibody

[0048] 1. Immunization of BALB / c mice

[0049] Preparation of rhNGF: Expressed, purified and prepared by the applicant (Le Wei, Peng Lujia, Shi Quanquan, etc. High-efficiency expression and biological activity evaluation of recombinant human nerve growth factor. China Pharmaceutical Affairs. 2014, 28(6): 601- 606.).

[0050] Select 5 female BALB / c mice, mix the rhNGF antigen with complete Freund's adjuvant (Sigma Company), and inject subcutaneously after complete emulsification. The immunization dose of each mouse is 80 μg for immunization. After the first immunization, the rhNGF antigen was emulsified with Freund's incomplete adjuvant (Sigma Company) at an interval of 2 weeks and 3 weeks, and then the immunization was repeated twice. Blood was collected from the tail of the mice, and the effect of serum-specific antibodies was detected by indirect ELISA method. Price (results such as figure 1 shown), and the ...

Embodiment 2

[0055] Example 2. Anti-hNGF Monoclonal Antibody Subclass Identification

[0056] The class and subclass of the monoclonal antibody were identified using the Rapid ELISA Mouse Antibody Isotyping Kit (Cat. No.: 37503) from Pierce, and the operation was performed according to the instructions of the kit.

[0057] Table 1 Identification results of anti-hNGF monoclonal antibody subtypes

[0058]

Embodiment 3

[0059] Example 3. Determination of the Affinity Constant of Anti-hNGF Monoclonal Antibody

[0060] Using the method of capturing antibodies, the affinity and binding kinetics of anti-hNGF monoclonal antibodies were detected with Biacore 3000, a biomolecular interaction analyzer from GE Company.

[0061] Table 2Biacore detection of the affinity of anti-hNGF monoclonal antibody and rhNGF antigen

[0062]

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Abstract

The invention relates to a neutralizing monoclonal antibody 12C11 of a human nerve growth factor and its hybridoma cell strain, which belong to the immunology field. The neutralizing monoclonal antibody 12C11 of the human nerve growth factor is secreted by the hybridoma cell with a preservation number of CGMCC No.8772, the neutralizing monoclonal antibody and the human nerve growth factor have neutralization reaction effect, and the subtype classification can be a heavy chain IgG1 and a light chain Kappa. The Neutralizing monoclonal antibody has specificity and neutralizing activity, and can be used as an antagonist for the human nerve growth factor.

Description

technical field [0001] The invention relates to the field of immunology, in particular to a monoclonal antibody against human nerve growth factor and a hybridoma cell line thereof. Background technique [0002] Nerve growth factor (NGF) is a nerve cell growth regulator with dual biological functions of neuron nutrition and neurite growth promotion. important regulatory role. [0003] Studies have shown that nerve growth factor (NGF) is a transmitter that plays a key role in the development of hypersensitivity and allodynia. A large number of preclinical studies have shown that blocking the interaction of NGF with its receptors can relieve pain. NGF monoclonal antibody neutralizes NGF and blocks the interaction between NGF and its receptors on sensory neurons to relieve pain. (Anti-nerve growth factor monoclonal antibody Tanezumab. Advances in Pharmaceutics. 2010,34(1):526.). Antibody antagonists of nerve growth factor have been reported to prevent or treat postoperative ...

Claims

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Application Information

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IPC IPC(8): C07K16/22C12N5/20A61K39/395A61P29/00A61P37/08A61P11/06A61P17/06G01N33/577C12R1/91
Inventor 刘荷中霍立红何景昌乐伟史权威
Owner 北京福睿君安科技有限公司
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