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Extraction, purification process for yersinia pestis natural F1 antigen

A Yersinia, F1 antigen technology, applied in the direction of bacterial antigen components, chemical instruments and methods, and resistance to vector-borne diseases, to achieve high immune protection, high sensitivity, and avoid the effects of protein activity

Inactive Publication Date: 2008-07-16
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The protein extracted by this method is of high purity, but organic solvents are still used, and the influence of organic solvents on proteins and environmental pollution cannot be avoided (Andrews GP, Heath DG, Anderson GW, Jr., Welkos SL, Friedlander AM: Fraction 1 capsularantigen (F1) purification from Yersinia pestis C092 and from an Escherichiacoli recombinant strain and efficacy against lethal plague challenge. Infect Immun 1996, 64(6): 2180-2187.)

Method used

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  • Extraction, purification process for yersinia pestis natural F1 antigen
  • Extraction, purification process for yersinia pestis natural F1 antigen

Examples

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Embodiment 1

[0034] Example 1, Extraction and Purification of Natural F1 Antigen of Yersinia pestis EV76 Vaccine Strain, Immunological Detection and Evaluation of Immunoprotective Effect

[0035] 1. Extraction and purification of natural F1 antigen of Yersinia pestis EV76 vaccine strain

[0036] Taking the Yersinia pestis EV76 vaccine strain as an example, the method of the present invention is used to extract and purify the natural F1 antigen from the Yersinia pestis EV76 vaccine strain. The specific process is as shown in Figure 1, including the following steps:

[0037] 1. Cultivation of Yersinia pestis EV76 vaccine strain

[0038] The Yersinia pestis EV76 vaccine strain (purchased from Lanzhou Institute of Biological Products) was inoculated into 5 mL of heart-brain infusion medium (purchased from BD Company, dissolve 37g of medium in 1L of pure water), culture at 37°C (35-40°C is acceptable) for 24h, then take 300μl of bacterial liquid, and then inoculate into 2L with 0.2% (0.1-0.3% ...

Embodiment 2

[0072] Example 2, Extraction and Purification of Natural F1 Antigen of Yersinia pestis A1122 Strain, Immunological Detection and Evaluation of Immunoprotective Effect

[0073] 1. Extraction and purification of natural F1 antigen of Yersinia pestis A1122 strain

[0074] Using the method of the present invention to extract and purify the natural F1 antigen from Yersinia pestis A1122 strain, the specific process is as shown in Figure 1, including the following steps:

[0075] 1. Cultivation of Yersinia pestis A1122 strain

[0076] Inoculate the Yersinia pestis A1122 strain (provided by the strain bank of the Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences) into 5 mL of heart-brain infusion medium supplemented with 0.1% (W / V, 0.1-0.3%) xylose (purchased from BD Company, 37g dissolved in 1L of pure water), cultured at 35°C (35-40°C is acceptable) and 250rpm for 12h, then took 300μl of bacterial liquid, and then inoculated into 2L with 0.3% (0.1-0.3...

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Abstract

The invention discloses an extraction and purification method for the natural F1 antigen of the Yersinia pestis, which adopts a proposal combining glass bead treatment, ammonium sulfate precipitation and molecular sieve filtration to successfully extract and purify natural F1 antigen with high purity from the Yersinia pestis cultures. The immunological detection and the animal protection experiment prove that the natural F1 antigen obtained by the method has higher protective immune function and detection sensitivity. If detecting the F1 monoclonal antibody of the same mouse ascites sample with the natural F1 antigen in the invention and the reformed F1 antigen (rF1) obtained from expression having the same concentration, the result indicates that the antibody titer detected by the natural F1 antigen is obviously higher than the detection result by the rF1. The extraction and purification method for the natural F1 antigen of the Yersinia pestis can play an important role in the development of Sub-unit Vaccine of plague and the detection and diagnosis of plague (e.g. immunological detection, etc.), which has a broad application prospect.

Description

technical field [0001] The invention belongs to the field of biology, and in particular relates to a method for extracting and purifying natural F1 antigen from the culture of Yersinia pestis. Background technique [0002] Plague is a severe infectious disease caused by Yersinia pestis. There are three types of human plague: bubonic plague, pneumonic plague, and septicemic plague. In recent years, the world's plague epidemic has been on the rise. In 2000, the World Health Organization listed plague as a re-emerging infectious disease (WHO: human plague in 1998 and 1999. weekly epidemiological record 2000, 75: 338-343.). In addition, Yersinia pestis is another [0003] The United States has listed Yersinia pestis as one of the six biological agents most likely to be used by terrorists. [0004] The inactivated plague vaccine only has a certain protective effect against bubonic plague, and the protection time is short, so this type of vaccine is not suitable for large-scale...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/24A61K39/02A61P31/04G01N33/569G01N33/53
CPCY02A50/30
Inventor 王效义王棠朱紫雯祁芝珍吴本传郭兆彪杨永海崔百忠王祖陨王虎杨瑞馥
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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