Unlock instant, AI-driven research and patent intelligence for your innovation.

A method for preparing monoclonal ascites antibody

A monoclonal, ascites technology, applied in the directions of biochemical equipment and methods, chemical instruments and methods, peptides, etc., can solve the problems of high requirements for production equipment and production environment, large antibody content, low ascites production, etc., to meet the feeding conditions The effect of low requirements, good experimental tolerance and strong reproductive ability

Active Publication Date: 2021-01-26
GUANGZHOU HUAYIN MEDICINE SCI & TECH
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Monoclonal antibodies are widely used in basic research and disease diagnosis in the biological field due to their high specificity; in recent years, with the development of science and technology, the problem of antibody heterogeneity has been solved and gradually used in the treatment of clinical diseases Very good effect; the amount of monoclonal antibody has increased rapidly, and the current method of preparing monoclonal antibody is high in cost and low in yield, which cannot meet the growing demand of the market
[0003] At present, there are two methods for preparing monoclonal antibodies. One is to use inbred BALB / c mice to prepare ascites antibodies. This strain of mice has good genetic consistency, small experimental individual differences, and large antibody content in ascites; the disadvantages are: This strain of mice has low fecundity and high requirements for feeding conditions, so the price is high. In addition, the mice are small in size and poorly tolerated by experiments, so the ascites production is low
The other is that with the development of cell fermentation technology, it has begun to be used for large-scale cultivation of monoclonal antibody hybridoma cells, but this method has extremely high requirements for production equipment and production environment, and the equipment price is also very expensive; The cost is high, and the requirements for technical and production personnel are also very high
Therefore, the price of antibody products is very expensive

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A method for preparing monoclonal ascites antibody
  • A method for preparing monoclonal ascites antibody
  • A method for preparing monoclonal ascites antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1) Hybridoma cell preparation: Influenza virus was cultured in chicken embryos, the allantoic fluid of chicken embryos was collected, the virus was purified, the spleen was taken after routine immunization of BALB / c mice, B lymphocytes were separated, fused with myeloma cells SP2 / 0, and in Culture in selection medium, and then clone by limited dilution method, select positive clones secreting antibodies for expansion culture, and harvest bone marrow hybridoma cells.

[0027] 2) Preparation of antibody ascites: divided into three groups: BALB / c mouse group, NIH mouse group and Kunming mouse group, 20 mice in each group, all 4 weeks old, BALB / c mice weighing 18 grams, NIH mice and The Kunming mouse group weighed 20 grams, intraperitoneally injected with 0.5ml of hypolipidane per mouse, and injected intraperitoneally with influenza virus hybridoma fusion cells on the third day, and the inoculation volume of each mouse was 1.0×10 6 cells

[0028] 3) Ascites collection: Asc...

Embodiment 2

[0039] 1) Preparation of hybridoma cells: Respiratory syncytial virus cultured with Hela cells, subjected to repeated freezing and thawing at low temperature, high-speed centrifugation to remove the precipitate, ultracentrifugation to remove the supernatant, and the precipitate was centrifuged with sucrose density gradient to obtain purified virus antigen; the antigen was routinely immunized with BALB Spleen was taken from / c mice, B lymphocytes were separated, fused with myeloma cells SP2 / 0, cultured in selective medium, and then cloned by limiting dilution method, and the positive clones secreting antibodies were selected for expansion culture, and harvested for bone marrow hybridization Tumor cells.

[0040] 2) Antibody ascites preparation: NIH mouse group and Kunming mouse group, 20 in each group, both 4-week-old NIH mice and Kunming mouse group, weighing 20 grams, intraperitoneally injecting 0.5ml of hypolipidane per mouse, on the third day Intraperitoneal injection of in...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for preparing monoclonal ascites antibody. The method is to inoculate outbred mice with hypolipidane intraperitoneally, inoculate myeloma hybridoma cells, and collect ascites every day from 3 to 7 days; the collected ascites is immediately Centrifuge with a centrifuge, place at 4°C, take it out the next day and then centrifuge and filter with filter paper, then use ammonium sulfate precipitation and column chromatography to purify to obtain the antibody. In the present invention, outbred mice are used instead of BALB / c mice to prepare monoclonal ascites antibodies. The advantages of this strain of mice are strong reproductive ability, low requirements on feeding conditions, and very cheap price, which is only small for BALB / c. It is about 1 / 5 of the mouse; and it is larger in size and has good tolerance to the experiment, so the output of ascites is large, which is several times that of the BALB / c mouse, and the antibody titer is not lower than that of the BALB / c mouse, and the antibody is sensitive to temperature The stability of BALB / c mice is much higher than that of BALB / c mice.

Description

technical field [0001] The invention relates to a method for preparing monoclonal ascites antibody. Background technique [0002] Monoclonal antibodies are widely used in basic research and disease diagnosis in the biological field due to their high specificity; in recent years, with the development of science and technology, the problem of antibody heterogeneity has been solved and gradually used in the treatment of clinical diseases Very good effect; the amount of monoclonal antibody has increased rapidly, but the current method for preparing monoclonal antibody is high in cost and low in yield and cannot meet the growing demand of the market. [0003] At present, there are two methods for preparing monoclonal antibodies. One is to use inbred BALB / c mice to prepare ascites antibodies. This strain of mice has good genetic consistency, small experimental individual differences, and large antibody content in ascites; the disadvantages are: This strain of mice has low fecundi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/00
CPCC07K16/00C07K2317/10
Inventor 李川江
Owner GUANGZHOU HUAYIN MEDICINE SCI & TECH