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A method and application of increasing monoclonal antibody production

A monoclonal antibody and yield technology, applied in chemical instruments and methods, biochemical equipment and methods, peptides, etc., can solve problems such as high cost and complicated process

Active Publication Date: 2021-07-30
ZYBIO INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, Wan Xiaochun and others injected the sensitizer twice to enhance the sensitization effect, thereby increasing the ascites production in mice [CN201210572740.3, Method for increasing the production of ascites-type monoclonal antibody, Wan Xiaochun, Lu Weidong, Yu Guang] , Hu Yumin and others increased the production of ascites in mice by adding Freund's incomplete adjuvant to the pristane (pristane) sensitizer [Hu Yumin, Ma Hongbing, Sun Zhongwen, etc., Freund's incomplete adjuvant Combined application of adjuvant and pristane in the preparation of ascites-type monoclonal antibody [J], SUZHOU UNIVERSITY JOURNAL OF MEDICAL SCIENCE 2003; 23(6)] However, the above methods all have problems such as high cost and complicated process, so there is an urgent need for a Method for increasing monoclonal antibody production with simple process and low cost

Method used

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  • A method and application of increasing monoclonal antibody production
  • A method and application of increasing monoclonal antibody production
  • A method and application of increasing monoclonal antibody production

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The screening process of parameter in the process of embodiment 1 present invention

[0044] 1. The screening of feeding glucose concentration is as follows:

[0045] (1) The anti-human NT-proBNP 6C7 hybridoma cells were cloned and cultured, and expanded in cell culture flasks. 10-14 days before the hybridoma cells were injected, 0.5ml / mouse of liquid paraffin was injected intraperitoneally into the mouse, and one week later, Dilute to 0.9×10 with PBS for injection 6 The hybridoma cells / ml were injected into the abdominal cavity of mice, 0.5ml / only, and a total of 70 mice were injected, divided into 7 groups;

[0046] (2) Feed the mice with pure water as the water source first, inject the cultured anti-human NT-proBNP 6C7 hybridoma cells into the abdominal cavity of the mice, and after 5 days, feed the mice in the 7 groups with a concentration of 0 %, 2%, 5%, 10%, 20%, 50%, 100% glucose solution, fed twice a day, fed with glucose until the antibody was collected;

[...

Embodiment 2

[0089] Embodiment 2 A preparation method of anti-human NT-proBNP monoclonal antibody 6C7, comprising the steps of:

[0090] (1) The anti-human NT-proBNP 6C7 hybridoma cells were cloned and cultured, and after being identified as all positive by ELISA, they were expanded in cell culture flasks, and one week before the intraperitoneal injection of the hybridoma cells, 0.5ml / head of liquid paraffin was injected for one week After injection, dilute to 0.9×10 with PBS 6 Hybridoma cells / ml were injected into the peritoneal cavity of mice, 0.5ml / only, and a total of 10 mice were injected;

[0091] (2) The mice were fed pure water all the time, and after the cultured anti-human NT-proBNP 6C7 hybridoma cells were injected into the abdominal cavity of the mice for 5 days, the mice were fed with 5% glucose, twice a day, and fed with Until the antibody collection is completed;

[0092] (3) 7-14 days after the cultured anti-human NT-proBNP 6C7 hybridoma cells were injected into the mice,...

Embodiment 3

[0098] The screening process of parameter in the process of embodiment 3 present invention

[0099] 1. The screening of feeding glucose concentration is as follows:

[0100] (1) Cloning and culturing the anti-human D-dimer 1D10 hybridoma cells, and expanding the culture in cell culture flasks. 10-14 days before injecting the hybridoma cells, intraperitoneally inject 0.5ml / mouse of liquid paraffin into the mouse, and one week later , diluted to 0.9×10 with PBS for injection 6 The hybridoma cells / ml were injected into the abdominal cavity of mice, 0.5ml / only, and a total of 70 mice were injected, divided into 7 groups;

[0101] (2) The mice were first fed with pure water as a water source, and the cultured anti-human D-dimer 1D10 hybridoma cells were injected into the abdominal cavity of the mice. After 5 days, the mice in the 7 groups were fed with a concentration of 0%, 2%, 5%, 10%, 20%, 50%, 100% glucose solution, fed once a day, fed with glucose until the antibody was coll...

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Abstract

The invention provides a method for increasing the production of monoclonal antibodies, comprising the following steps: (1) cloning and culturing hybridoma cells, and injecting the cultured hybridoma cells into animals; (2) feeding glucose solution to animals (3) collecting ascites antibody; (4) purifying the collected ascites antibody. In the present invention, after the animals are injected into the hybridoma cells, the mice start to be fed with the glucose solution mixed with pure water, and unexpected technical effects are obtained. The monoclonal antibody obtained by the method is equivalent, greatly reduces the cost of antibody production, greatly increases its economic benefits, and provides a new idea for the supply of raw materials in the IVD industry.

Description

technical field [0001] The present invention relates to an antibody preparation method and its application, in particular to a method for increasing monoclonal antibody production and its application. Background technique [0002] As a popular research object in the pharmaceutical industry, monoclonal antibodies are widely used in various biological related fields. In this environment, technology platforms such as immunoturbidimetry, immunochromatography, and chemiluminescence, which use antibodies as the core raw materials, have also made great progress. However, the antibody raw materials used in it have problems of insufficient yield and uneven quality, which has become a key link restricting the quality of domestic reagents. Therefore, it is particularly important to improve the production of antibodies and control the quality. The preparation methods of monoclonal antibodies include in vitro cell culture method and animal in vivo induction method. The in vitro culture...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/00C12N5/20
CPCC07K16/00C07K2317/14
Inventor 郭绮代玉兰任燕斌
Owner ZYBIO INC
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