Preparation method and application of bacillus cereus monoclonal antibody

A Bacillus cereus, monoclonal antibody technology, applied in the biological field, can solve the problems of many influencing factors, time-consuming, poor sensitivity, etc., to achieve the effect of promoting animal immunity, ensuring animal welfare, and good specificity

Inactive Publication Date: 2016-01-13
FUDEAN OF BEIJING TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Although the routine detection method of Bacillus cereus is easy to operate, there are too many influencing factors, time-consuming, and poor sensitivity.

Method used

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  • Preparation method and application of bacillus cereus monoclonal antibody
  • Preparation method and application of bacillus cereus monoclonal antibody
  • Preparation method and application of bacillus cereus monoclonal antibody

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Embodiment Construction

[0032] specific implementation plan

[0033] The present invention is further described in detail by the following examples.

[0034] Preparation of Whole Cell Antigen of Bacillus cereus

[0035] Resuscitation and activation of Bacillus cereus in this laboratory. Prepare LB medium (weigh 10 g of tryptone, 5 g of yeast yeast powder and 10 g of NaCl per liter, pH 7.0), and autoclave at 121° C. for 20 minutes. Under sterile environment conditions, add 1mL of Bacillus cereus preserved at ~80°C to 25mL of liquid medium, and culture in an incubator at 37°C and 220rpm for 15h. Centrifuge at 10,000rpm for 25min and discard the supernatant to obtain bacterial pellet. Prepare sterile physiological saline, weigh 0.9g NaCl and dissolve in 100mL distilled water, and autoclave at 121°C for 20 minutes. Resuspend the above bacterial pellet with sterile normal saline, centrifuge at 10,000rpm for 20min, pour off the supernatant, wash 3 times in reciprocating cycles, and resuspend with 4% pa...

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Abstract

The invention provides a preparation method and application of a bacillus cereus monoclonal antibody. The preparation method comprises the following steps of: 1, preparing bacterial antigen through fixation of bacillus cereus by incubation treatment of paraform; 2, injecting the bacterial antigen so as to avoid emulsification; 3, performing short spacing impulse injection so as to quickly promote immunization; 4, planting solid tumors so as to rejuvenate tumor cells; 5, performing chemical induction and fusion; 6, performing semifluid selection so as to obtain monoclonal hybridoma; 7, performing cooperation of negative screening and positive screening so as to obtain a specific antibody; and 8, using a polyclonal antibody as a capture antibody and using a monoclonal antibody as a detection antibody to establish dual-antibody sandwich ELISA.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a preparation method and application of a Bacillus cereus monoclonal antibody. Background technique [0002] Food poisoning is a food-borne disease that seriously endangers human health. Most food poisoning is caused by harmful bacteria contaminating food. In bacterial food poisoning, Bacillus cereus food poisoning is more common. Bacillus cereus is an aerobic and facultative anaerobic Gram-positive bacillus widely found in a variety of plant foods and raw and cooked foods. Bacillus cereus is an opportunistic pathogenic bacterium that can produce diarrhea toxin and vomitoxin to cause food poisoning. Sometimes it can also cause human eye diseases, endocarditis, meningitis and other diseases through bacterial infection. As early as 1906, the first Bacillus cereus food poisoning was reported, and then many northern and eastern European countries also reported similar poisonings. Accor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12G01N33/577G01N33/569
Inventor 罗云波许文涛朱龙佼黄昆仑邵向丽解沛燕徐瑗聪翟百强
Owner FUDEAN OF BEIJING TECH
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