Preparation method of bacillus cereus polyclonal antibodies

A technology of Bacillus cereus and polyclonal antibody, applied in the direction of anti-bacterial immunoglobulin and immunoglobulin from serum, etc., can solve the problems of time-consuming, poor sensitivity, and many influencing factors, and achieve strong sensitivity and stimulate immunity Response, high potency effect

Inactive Publication Date: 2014-05-21
FUDEAN OF BEIJING TECH +1
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Problems solved by technology

[0006] Traditional serological reactions such as one-way / two-way diffusion technology, indirect hemagglutination technology, convective immunological agarose gel electrophoresis technology, complement fixation test, etc. are easy to operate, but there are too many influencing factors, time-consuming, and poor sensitivity.

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  • Preparation method of bacillus cereus polyclonal antibodies
  • Preparation method of bacillus cereus polyclonal antibodies
  • Preparation method of bacillus cereus polyclonal antibodies

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[0031] specific implementation plan

[0032] The present invention is further described in detail by the following examples.

[0033] Preparation of Bacillus cereus antigen

[0034]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. Prepare solid LB medium (weigh 10g tryptone, 5g yeast powder, 10g NaCl and 15g agar powder per liter, pH 7.0), autoclave at 121°C for 20 minutes, when the temperature drops to ~50°C, In a sterile environment, pour the culture medium into an autoclaved plate and let it solidify. Take 200 μL of the activated bacteria from the above liquid culture and spread evenly on the solidified medium, and culture at 37°C for 15 hours. ...

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Abstract

The invention provides a preparation method of bacillus cereus polyclonal antibodies, which comprises the following steps: 1, enrichment: coating activated bacillus cereus on a flat plate for enlarge cultivation; 2, fixation: fixing surface compositions of the bacillus cereus by using a modified boiling water bath method, so that antigenic determinants are fully exposed; 3, emulsification: preparing a uniform emulsified liquid from an adjuvant-bacillus mixture by using a self-assembled emulsifier; 4, immunization: carrying out animal immunization in a mode of being based on multiple loca on the back and supplemented by peripheral loca; 5, antiserum titer: evaluating the antiserum titer by using an ELISA method, and carrying out enhanced immunization; 6, taking whole blood: taking whole blood from cervical arteries under the condition of anesthesia by using chloral hydrate; 7, preparation of antiserum: standing the rabbit whole blood at optimum temperature so as to carry out natural chromatography, and carrying out centrifugal separation on the antiserum; and 8, purification: purifying the antiserum by using an n-caprylic acid-ammonium sulfate method, so that a polyclonal antibody is prepared. The polyclonal antibody prepared by using the preparation method disclosed by the invention is high in titer, good in specificity and strong in sensitivity, and has a great application advantage.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of a polyclonal antibody against Bacillus cereus. Background technique [0002] The first cases of Bacillus cereus food poisoning were reported as early as 1906. However, the etiology was not clarified until 1950, after which many countries, especially northern and eastern European countries, also reported similar poisonings. Bacillus cereus was the third most common pathogen in bacterial food poisoning in Hungary from 1960 to 1968. The food poisoning caused by this bacteria accounted for 84% of the reported cases of food poisoning, and the number of poisoned people accounted for the total number of poisoned people. 14.8%. Since 1971, there have been many reports of food poisoning caused by this bacteria in the UK. In 1973, the Nanjing Municipal Health and Epidemic Prevention Station reported for the first time that children in a nursery in Nanjing...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12C07K16/06
Inventor 许文涛朱龙佼黄昆仑何景罗云波翟百强董凯
Owner FUDEAN OF BEIJING TECH
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