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
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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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