Pseudomonas aeruginosa resisting Fab' fragment

A technology of Pseudomonas aeruginosa and fragments, applied in the field of immunoglobulin, can solve the problems of poor therapeutic effect and achieve stable physical and chemical properties, high yield and good potency

Inactive Publication Date: 2010-09-01
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the antibiotics used for Pseudomonas aeruginosa infection are constantly updated, due to the broad-spectrum drug resistance of Pseudomonas aeruginosa, the treatment effect is often poor, making it an urgent need to find alternative treatments for antibiotics

Method used

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  • Pseudomonas aeruginosa resisting Fab' fragment
  • Pseudomonas aeruginosa resisting Fab' fragment
  • Pseudomonas aeruginosa resisting Fab' fragment

Examples

Experimental program
Comparison scheme
Effect test

example 2

[0039] Example 2 Identification of anti-Pseudomonas aeruginosa Fab' fragment of the present invention

[0040] 1. The highest positive titer of anti-Pseudomonas aeruginosa Fab' detected by indirect ELISA is 1:25600.

[0041] The anti-Pseudomonas aeruginosa Fab' was diluted to 1 / 400, 1 / 1600, 1 / 6400, 1 / 25600, 1 / 51200, 1 / 102400, and its absorbance value at 450nm was detected by indirect ELISA reaction. The specific results are as follows figure 1 As shown, take the logarithmic value of the dilution factor as the horizontal axis, and draw a smooth curve with the absorbance as the vertical axis. It can be seen that the change of absorbance and the dilution factor basically form a slanted line). Positive judgment of the titer of the anti-endotoxin Fab' detected by indirect ELISA: (measured OD value-blank control OD value) / (negative control OD value-blank control OD value)>2.1. The negative control was unimmunized chicken Fab' (1:100). According to the positive judgment standard, t...

example 1

[0042] Example 1 Preparation of anti-Pseudomonas aeruginosa Fab' fragment of the present invention

[0043] The anti-Pseudomonas aeruginosa Fab' fragment of the present invention is extracted from eggs of laying hens immunized with Pseudomonas aeruginosa as an antigen, and the specific method is:

[0044] (1) Pseudomonas aeruginosa (ATCC27853, the standard strain preserved in the Laboratory Department of Southwest Hospital) was purely cultured and adjusted to 1.5×108 / ml bacterial liquid, then ultrasonically crushed and used as an immunogen for future use. The 25-week-old Leghorn chickens were immunized after one week of routine feeding. The immunogen supplemented with complete Freund's adjuvant was injected subcutaneously in both wings and in the muscles of both thighs for immunization. After the first immunization, booster immunization was given once every two weeks, for a total of 4 immunizations. Eggs were collected 2 weeks after the first immunization and stored for later...

example 3

[0066] Example 3 The in vitro biological effect experiment of the anti-Pseudomonas aeruginosa Fab' fragment of the present invention

[0067] 1. Experimental method:

[0068] The experiment was divided into two groups, one of which was the control group: 3.9×105cfu / ml Pseudomonas aeruginosa liquid was incubated in a 37°C incubator, and the viable bacteria were counted after sampling at 6, 12, 24, and 36 hours. Another group is the Fab' fragment antagonism group of the present invention: add Pseudomonas aeruginosa (3.9 * 105cfu / ml) into Fab' fragment 5, 10, 20mg / ml of the present invention respectively, at 6, 12, 24, 36h, live after sampling bacteria count.

[0069] 2. Experimental results: if image 3 As shown, the anti-Pseudomonas aeruginosa Fab' can obviously inhibit the growth of Pseudomonas aeruginosa, and has a good biological effect in vitro.

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Abstract

The invention provides a specific antibody of bacillus pyocyaneus, in particular to a bacillus pyocyaneus resistant Fab' fragment, which is extracted from the egg of laying hens immunized via bacillus pyocyaneus antigen, with high yield, better effect, stable physicochemical property, high specificity, simple preparation, better repeatability, and industrialization production support. The inventive bacillus pyocyaneus resistant Fab' fragment can significantly resist the growth of bacillus pyocyaneus, by external measurement, which can be used to prepare the drug treating bacillus pyocyaneus resistant infection.

Description

technical field [0001] The present invention relates to an immunoglobulin, in particular to a Fab' fragment of an immunoglobulin. Background technique [0002] Pseudomonas aeruginosa is the most common opportunistic pathogen in the genus Pseudomonas. It is widely distributed in nature and can be transmitted through environmental pollution, cross-infection, endogenous infection and iatrogenic infection. Pseudomonas aeruginosa has stronger resistance to the outside world than other bacteria, can survive for a long time in humid places, has strong resistance to dryness, and has low nutritional requirements. Because of its multi-drug resistance and other characteristics, it has become the main pathogenic bacteria in hospital infections. Pseudomonas aeruginosa infection is closely related to burns, low immune function, sepsis, lower respiratory tract infection, etc., and the fatality rate is extremely high. Although the antibiotics used for Pseudomonas aeruginosa infection are c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/02A61K39/40A61P31/04
Inventor 张雅萍庄颖马思远
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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