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Anti-human IgG monoclonal antibody, hybridoma cell line and application thereof

A hybridoma cell line, monoclonal antibody technology, applied in anti-viral immunoglobulin, biochemical equipment and methods, instruments and other directions, can solve the problems of reducing reliability, rising costs, etc. Effects of relaxed storage and handling conditions

Active Publication Date: 2019-01-25
SICHUAN ANKERUI NEW MATERIAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, antibody titer, cross-reactivity, and stability are all important evaluation factors. For example, the level of antibody titer reflects the minimum titer of reacting with the antigen at a certain concentration, and the lower the titer, the higher the titer; antibody cross-reactivity The stability of the antibody can affect the specificity of the antibody; the stability of the antibody will directly affect the reliability of the final result, and the antibody with poor stability requires higher storage conditions and operating conditions, thereby reducing its practicability and effectiveness as a diagnostic reagent. The reliability of the results, and inevitably lead to rising costs; clinical trials are used to illustrate the actual effect of antibodies in diagnosing corresponding diseases or viral infections

Method used

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  • Anti-human IgG monoclonal antibody, hybridoma cell line and application thereof
  • Anti-human IgG monoclonal antibody, hybridoma cell line and application thereof
  • Anti-human IgG monoclonal antibody, hybridoma cell line and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] The immunization of embodiment 1 mice

[0062] Human blood-derived IgG antigen (Sichuan Mike Bio-New Material Technology Co., Ltd., batch number 051519) was diluted to 3.0 mg / ml with normal saline, and mixed with an equal volume of Freund's complete adjuvant (Sigma Company, product number SLBF-9338V) (50 μg / ml). BALB / c mice), emulsified into an oily emulsion with a 1ml syringe, until the oily emulsion dripped into the water does not disperse and then stop the emulsification, the emulsion is subcutaneously administered to the BALB / c mice with a dose of 100 μl / axillary limbs ( Chengdu Dashuo Experimental Animal Center, 5-week-old females, 4) Immunization was enhanced 7 days after the first immunization, and human IgG (Sichuan Mike Bio-New Material Technology Co., Ltd., batch number 051519) and Freund's incomplete adjuvant (Sigma Company) were taken to enhance immunity. , Cat. No. SLBM9367V) equal volume mixed (25μg / mouse BALB / c) and then emulsified, the immunization dose ...

Embodiment 2

[0065] The preparation of embodiment 2 hybridoma cell lines

[0066] 2-1 Preparation of feeder cells

[0067] Peritoneal macrophages of normal 12-week-old BALB / c mice were used as feeder cells. One day before the fusion, BALB / c was sacrificed by taking blood from the eyes and pulling the neck, soaking in 0.1% bromogeramine for 1 minute, then soaking in 75% alcohol for 1 minute, lifting the abdominal skin from the hind abdomen with sterile scissors in an ultra-clean bench to expose the peritoneum . Wipe the peritoneum with an alcohol swab to disinfect. Inject 2ml of RPMI1640 culture solution into the abdominal cavity with a syringe, taking care to avoid penetrating into the intestine. Fix the syringe with the right hand so that the needle remains in the abdominal cavity, and gently massage the abdomen with the alcohol cotton ball in the left hand for 1 minute, and then suck out the injected culture solution. Centrifuge at 1000r / min for 5-10 minutes, discard the supernatant....

Embodiment 3

[0080] The preparation of embodiment 3 monoclonal antibody

[0081] Select healthy BALB / c mice of 12-14 weeks, inject 0.5ml liquid paraffin (Tianjin Kemiou) into each mouse intraperitoneally, and inject 1.8×10 6 a hybridoma cell. Ascites can be produced 7-10 days after cell inoculation. Observe the occurrence of ascites in mice every day. If the abdomen is obviously enlarged and the skin feels tense when touched with hands, the mice can be killed by pulling the neck, and the ascites can be sucked into the test tube with a dropper. One mouse can obtain 1-5ml ascites. The collected ascites was centrifuged to obtain the supernatant, and a small sample was taken and stored in a -20°C refrigerator. The ascitic fluid was saturated and precipitated with ammonium sulfate, and then purified with a protein A affinity chromatography column, and the purity of the antibody (denoted as human IgG-Ab) detected by SDS-PAGE was greater than 90%.

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Abstract

The invention relates to a hybridoma cell stain and a monoclonal antibody secreted by the hybridoma cell strain, and the antibody can specifically bind to human IgG. The invention also relates to a kit comprising the hybridoma cell strain or monoclonal antibody. The monoclonal antibody of the invention exhibits good performance in the aspects of antibody purity, repeatability, antibody titer and stability.

Description

technical field [0001] The present invention relates to a monoclonal antibody, in particular to an anti-human IgG monoclonal antibody, a hybridoma cell secreting the monoclonal antibody, and the application of the monoclonal antibody. Background technique [0002] Herpes simplex virus (HSV) is a kind of virus with envelope and double-stranded DNA genome. It exists widely in nature and can infect humans and many animals. It has strong tropism especially for human skin tissue. , is a common pathogen of skin diseases and venereal diseases. There are two serotypes of herpes simplex virus, including herpes simplex virus-1 (HSV-1) and herpes simplex virus-2 (HSV-2). Genital herpes caused by HSV-1 and HSV-2 has caused more and more serious social and public health problems. [0003] At present, the commonly used laboratory methods for detecting HSV include direct immunofluorescence or histochemical staining, virus isolation and culture, antigen latex agglutination test, serum ant...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/08G01N33/577G01N33/569C12R1/91
CPCG01N33/56994G01N33/577C07K16/087G01N2333/035
Inventor 舒川黄家菊李岚敏王磊
Owner SICHUAN ANKERUI NEW MATERIAL CO LTD
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