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Nicotinamide N-methytransferase (NNMT) protein monoclonal antibody, hybridoma cell strain and application

A hybridoma cell line, monoclonal antibody technology, applied in the direction of anti-enzyme immunoglobulin, biochemical equipment and methods, instruments, etc., can solve the problems affecting the effect of radiation and radiotherapy, and achieve the effect of high antibody titer

Inactive Publication Date: 2012-08-01
ZHEJIANG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These studies suggest that NNMT is closely related to tumorigenesis. It is speculated that NNMT may participate in various biological functions of tumor cells, be related to the occurrence and development of tumors, and may affect the effect of Chinese medicine and radiotherapy.

Method used

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  • Nicotinamide N-methytransferase (NNMT) protein monoclonal antibody, hybridoma cell strain and application
  • Nicotinamide N-methytransferase (NNMT) protein monoclonal antibody, hybridoma cell strain and application
  • Nicotinamide N-methytransferase (NNMT) protein monoclonal antibody, hybridoma cell strain and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Obtaining of hybridoma cell lines, screening and preparation of monoclonal antibodies

[0031] 1. Preparation of monoclonal antibodies

[0032] 1.1. Preparation of immunogen

[0033] The immunogen is GST-NNMT fusion protein and NNMT protein, and the NNMT protein is prepared by genetic engineering technology: the pGEX-4T-2 / NNMT plasmid is constructed and transformed into E. coli BL-21-STAR (DE3) to express the GST-NNMT fusion protein . Then, after the fusion protein was purified, the fusion protein was fully digested by adding thrombin, and the NNMT protein (geneBank accession number: ACA06031.1) was obtained after purification again.

[0034] 1.2. Animal immunity:

[0035] The immunization procedure is as follows: for the first immunization, take 6-8 week-old BALB / C female mice, mix NNMT protein and Freund's complete adjuvant in equal volumes, and then inject subcutaneously at multiple points on the back of the mice, with a dose of 120 μg per mouse . Aft...

Embodiment 2

[0082] Example 2: Application of monoclonal antibody to immunohistochemical detection of clinical specimens (two-step indirect method)

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Abstract

The invention provides a hybridoma cell strain of NNMT protein monoclonal antibody, a monoclonal antibody secreted by the cell strain and application. The monoclonal antibody of the invention has a high antibody titer, affinity and high purity after purification to NNMT proteins, thus indicating that the monoclonal antibody can be applied in the scientific research field of NNMT proteins as well as in clinic detection for immunohistochemistry of pathological sections of different pathologic tissues to detect expression of NNMT proteins in the tissues. Therefore, the monoclonal antibody provided in the invention contributes to diagnosis, treatment and prognosis of NNMT associated tumors.

Description

(1) Technical field [0001] The present invention relates to nicotinamide-N-methylase (NNMT, EC2.1.1.1) protein monoclonal antibody hybridoma cell line, monoclonal antibody, and monoclonal antibody in preparation and detection of nicotinamide-N-methylase Application of protein kits. (2) Background technology [0002] Malignant tumors seriously endanger human health. According to the latest data from the World Health Organization, by 2020, there will be 15 million new cancer patients every year. Not only that, the number of cancer deaths is also rising rapidly around the world. In 2007, a total of 7.6 million people died worldwide. For cancer, this number may increase to 13.2 million in 2030. [0003] Early diagnosis of tumors is the key to cancer prevention and treatment. The risk of death and recurrence of tumors is closely related to the stage at the time of initial diagnosis, and the sensitivity of chemotherapy and radiotherapy depends to a certain extent on the enzymes o...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/40G01N33/577C12R1/91
Inventor 张钧谢鑫友杨肃文王燕忠
Owner ZHEJIANG UNIV
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