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Method for preparing anti-duck plague transfer factor

A technology of transfer factor and duck plague, which is applied in peptide preparation methods, chemical instruments and methods, antiviral agents, etc., can solve the problems of incomplete cell fragmentation, large loss of product activity, and difficulty in improving the yield, and achieves improved Effects of cure rate, reduction of morbidity rate, and easy source

Inactive Publication Date: 2008-10-29
TIANJIN SHENGJI GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The content of the patent application that has been reported is to directly filter and extract the broken tissue after freezing and centrifuging, which also has the defect of incomplete cell crushing, and the process is cumbersome, the time is long, and the activity loss of the product is also large. Therefore, It is difficult to increase the yield
[0006] Porcine spleen and thymus are places where immunocompetent cells gather, and the sources of materials are very extensive. Therefore, it is of great theoretical and practical significance to extract specific transfer factors from these cells, but it has not been reported yet.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Preparation of Anti-Duck Plague Transfer Factor

[0029] 1. Preparation of virus antigen: Inoculate duck plague into the allantoic fluid of 10-day-old duck embryos, 48-96 hours at 33°C, take the allantoic fluid of duck embryos, put it in a small thick-walled bottle, crush it by ultrasonic, and centrifuge at 5000rpm for 20 minutes , discard the sediment and leave the supernatant;

[0030] 2. Take the centrifuged supernatant, then discontinuous gradient density centrifugation and purification to extract the viral antigen; after the viral antigen is centrifuged, take the supernatant and fill it with discontinuous gradient density prepared with 15%, 30%, 45% and 60% sucrose Centrifuge tube, ultracentrifuge, 165,000 rpm for 3 hours, determine the position of the centrifuge tube where the virus is located according to the molecular weight of different viruses, and suck it out for later use;

[0031] 3. Immune pigs with the above-mentioned extracted virus antigens: use the ab...

Embodiment 2

[0042] Preparation of Anti-Duck Plague Transfer Factor

[0043] (1) Preparation of raw materials: 10 healthy 6-month-old pigs with no medical history were selected and immunized with duck plague vaccine subcutaneously. After 15 days, they were immunized again with the same method as above. Slaughter 15-20 days after the second immunization, take the spleen and thymus on ice, transfer and store at -20°C for later use;

[0044] (2) Pretreatment: After weighing the pig spleen, wash the pig spleen with cold three-distilled water, cut off its fascia and adipose tissue with scissors, and then wash it with cold three-distilled water;

[0045] (3) Crushing: Cut the pig spleen washed above into pieces, add 2 times the volume of cold normal saline, and mash it with a high-speed tissue masher (1000r / min) for 3 times under freezing conditions, 3 minutes each time, to prepare get homogenate;

[0046] (4) Freezing and thawing: place the homogenate (packed in a container) in an ultra-low t...

Embodiment 3

[0053] Detection of the anti-duck plague transfer factor prepared by the process of the present invention

[0054] The anti-duck plague transfer factor (hereinafter referred to as "this product") prepared by the process described in Example 1 is detected as follows:

[0055] (1) Ultraviolet spectrophotometric measurement: This product has a high absorption peak at 250.0-252.0nm, and ABS260 / ABS280>2.0.

[0056] (2) The standard solution is light yellow and the pH value is between 6.0-6.5.

[0057] (3) 20% sulfosalicylic acid test: This product has no turbidity and precipitation, indicating that the protein reaction is negative, and it does not contain macromolecular proteins.

[0058] (4) Determination of peptide content: The peptide content of this product was determined to be 1.130 mg / ml by the biuret method.

[0059] (5) Determination of nucleic acid content: The nucleic acid content of this product was determined to be 602.78ug / ml by the orcinol method.

[0060] (6) Bact...

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PUM

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Abstract

The invention discloses a preparation method of a natural high-effective biological active substance anti-duck plague virus transfer factor. The preparation method comprises the following steps of: inoculating duck plague virus into an allantoic fluid obtained from a 10-day-aged chick embryo, and preparing duck plague virus antigen; extracting the duck plague virus antigen; immunizing a pig with the extracted duck plague virus antigen; and extracting the anti-duck plague virus transfer factor from the immunized pig. The inventive anti-duck plague virus transfer factor can prevent duck plague and protect normal body cells from being infected by the duck plague virus, so as to reduce incidence rate.

Description

technical field [0001] The invention relates to the field of medicines and preparations thereof, in particular to a preparation method of anti-duck plague transfer factor. Background technique [0002] Duck plague is an infectious disease caused by duck plague virus, and occurs mostly in ducks after 20 days of age. The main symptoms are: elevated body temperature, dyspnea, tearing, green loose stools, swelling of the head and neck of the sick duck, so it is called "big head plague". The morbidity and mortality of duck plague are high [0003] Although there are some medicines or preparations against duck plague at present: but because the pertinence of existing medicine preparations is not strong, curative effect is limited, moreover, therapeutic preparations are just applied after the onset mostly, can not play preventive effect. For duck plague-susceptible duck flocks, regular vaccination of duck plague every year is an effective way to prevent duck plague. Secondly, nu...

Claims

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Application Information

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IPC IPC(8): C07K1/14C07K1/34A61P31/14
Inventor 田杏芳王连民
Owner TIANJIN SHENGJI GRP CO LTD
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