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

A hybridoma cell line and monoclonal antibody technology, applied in the field of medical testing, can solve the complex problems of monoclonal antibodies, achieve the best immune effect, excellent long-term and thermal stability, and prolong the service life

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

AI Technical Summary

Problems solved by technology

[0004]However, it is very complicated to obtain monoclonal antibodies that can be used in in vitro diagnostic kits. First, it is necessary to screen high-purity and active antigens to prepare enough Antibodies, then systematically evaluate the antibodies to obtain clinically relevant candidate antibodies, and then develop them into detection reagents

Method used

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

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Experimental program
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Effect test

Embodiment 1

[0061] The immunization of embodiment 1 mice

[0062] Human blood-derived IgG antigen (Sichuan Mike New Material Technology Co., Ltd., batch number 071516) was diluted to 2.0 mg / ml with normal saline, and mixed with an equal volume of Freund's complete adjuvant (Sigma Company, product number SLBF-9338V) (100 μg / piece BALB / c mouse), be emulsified into oily emulsion with 1ml syringe, stop emulsification until the oily emulsion dripped into water does not disperse, and this emulsion is subcutaneously administered to BALB / c mouse (Chengdu Dashuo Experimental Animal Center, 5-week-old female, 2) Immunization was enhanced 14 days after the first immunization, human IgG was mixed with an equal volume of Freund's incomplete adjuvant (Sigma Company, product number SLBM9367V) (50 μg / body BALB / c After emulsification of mice), the immunization dose was 50 μl / mouse, and the immunization was boosted once every other week, and the tail blood was collected before each immunization, and the se...

Embodiment 2

[0066] The preparation of embodiment 2 hybridoma cell lines

[0067] 2-1 Preparation of feeder cells

[0068] 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 2 mL 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

[0081] The preparation of embodiment 3 monoclonal antibody

[0082] Select healthy BALB / c mice of 12-14 weeks, inject 0.5mL liquid paraffin (Tianjin Kemiou) intraperitoneally into each mouse, and inject 2×10 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 ascites was saturated and precipitated with ammonium sulfate, and then purified with a protein A affinity chromatography column. The purity of the antibody (denoted as human IgG-Ab1) detected by SDS-PAGE was greater than 90%, and the results were as follows: figure 1 shown.

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Abstract

The invention discloses an anti-toxoplasma gondii IgG monoclonal antibody, a hybridoma cell strain, a toxoplasma gondii detection kit and application thereof. The hybridoma cell strain is preserved inthe China Center for Type Culture Collection, and the preservation number is CCTCC NO: C2018175. The monoclonal antibody which is secreted by the hybridoma cell disclosed by the invention is specifically bound with an anti-toxoplasma gondii IgG antibody. The hybridoma cell strain and the monoclonal antibody which are provided by the invention have the advantages of high specificity, high sensitivity, good stability and no cross reaction, and can be applied to the kit of the IgG antibody for detecting toxoplasma gondii infection.

Description

technical field [0001] The invention belongs to the field of medical testing, and in particular relates to an anti-human IgG monoclonal antibody, a hybridoma cell line, a kit for detecting toxoplasma gondii and an application thereof. Background technique [0002] Toxoplasma gondii is an opportunistic pathogenic protozoan that can cause toxoplasmosis in humans and animals. Toxoplasma gondii infection in adults is mostly recessive, only showing positive serum toxoplasma-specific antibodies. When the immune function is impaired, it can turn into acute toxoplasmosis, causing headaches, coma, and sudden vision loss. The consequences are very serious. The disease is distributed worldwide, and the infection rate of the normal population in my country is mostly below 10%, with an average infection rate of 5.17%. Toxoplasma gondii can be transmitted through blood transfusion, causing harm to recipients of blood transfusion, especially to pregnant women, which can cause fetal malfor...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/42G01N33/577G01N33/569
CPCG01N33/56905G01N33/577C07K16/425C07K2317/35C07K2317/33C07K2317/94
Inventor 舒川黄家菊李岚敏王磊
Owner SICHUAN ANKERUI NEW MATERIAL CO LTD
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