Method for preparing anti-dog parvovirus egg yolk antibody biological preparation

A technology of canine parvovirus and biological preparations, applied in antiviral agents, antiviral immunoglobulins, antibodies, etc., can solve the problems of high cost, large loss of IgY activity, cumbersome preparation process, etc., and achieve low cost, quick effect, The effect of preventing puppies from parvovirus infection

Inactive Publication Date: 2007-10-31
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method requires multi-step purification of the yolk antibody before use, the preparation process is cumbersome, the cost is high, and the activity of IgY is greatly lost

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Canine parvovirus field virus collection: take the intestines and contents of dead dogs diagnosed as parvovirus infection, add normal saline to grind at a weight ratio of 1:1, centrifuge to remove sediment, and supernatant 20000rpm high-speed centrifugation for 60 minutes to remove tiny particles.

[0016] Initial virus extraction: add chloroform to 10% of the total supernatant, shake vigorously, and centrifuge to remove fat-soluble substances.

[0017] Virus adsorption: 10% of the total supernatant was added to washed porcine red blood cells hydroformylated with 2.5% glutaraldehyde by volume, shaken well and placed at 4°C for 2 hours, and centrifuged to precipitate porcine red blood cells.

[0018] Virus release: use 4% hypertonic saline to make the precipitated porcine red blood cells into a suspension with a volume ratio of 10%, centrifuge at 37° C. for 2 hours to remove the porcine red blood cells.

[0019] Determination of HA titer: the virus titer in the released ...

Embodiment 2

[0028] Canine parvovirus field virus collection, virus initial extraction, virus adsorption and release test purification, HA value detection, and vaccine preparation are the same as in Example 1. For the first immunization, the immunization dose is 0.5ml per head. The second immunization was carried out at an interval of 10 days, and the immunization dose was 1 ml. After 14 days of immunization, the titer of egg yolk was measured by hemagglutination inhibition test. When the titer was lower than 1:128, the third immunization was carried out, and the immune dose was 1ml. After 14 days, the immunized eggs were collected, and the anti-parvovirus egg yolk antibody The preparation method of the biological preparation is the same as in Example 1.

[0029] Preparation of vaccine, immunization, IgY extraction and purification by the above method, the yield of IgY is over 95%, and the activity is not affected in any way. More than 26 mg of IgY can be obtained after extraction from 5 ...

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Abstract

The invention discloses a preparing method of anti-dog parvoviral vitelline antibody biological preparation, which comprises the following steps: collecting disease material of dead dog with parvoviral infection; centrifuging; removing depositing slag; centrifuging the supernatant fluid; adding into chloroform; vibrating severely; centrifuging; getting the supernatant fluid; aldehyde-washing pig red cell; adding the supernatant into the pig red cell; vibrating evenly; stewing; centrifuging; getting depositing liquid; adding into salt water; stewing; centrifuging; getting virus liberating liquid; testing HA potency with liberating liquid; reaching 1: 4096 effective; deactivating the virus liberating liquid with methyl aldehyde; adding into oil emulsion dog parvoviral vaccine prepared by brown algae polysaccharide; passing the vaccine to hen after delivery; testing HI potency of immune ovum with hemagglutination inhibition test; harvesting vitelline after reaching standard; diluting with sterile distilled water; proceeding acid treatment with lactic acid liquid; removing mixed protein; degreasing with freeze-thaw; salting-out suction liquid; adjusting IgY density; loading; putting into bathing box; treating; getting the oral biological preparation.

Description

technical field [0001] The invention relates to a preparation method of a biological preparation, in particular to a preparation method of an anti-canine parvovirus yolk antibody biological preparation. used in the field of bioengineering. Background technique [0002] Canine parvovirus (CPV) enteritis is one of the most common infectious diseases in dogs, and its morbidity and mortality are as high as 50% to 100%, seriously affecting the health of dogs. In recent years, with the increase in the number of meat dogs and pet dogs in urban and rural areas, the incidence of the disease is on the rise. At present, symptomatic and supportive treatments are generally used for sick dogs, and the cure rate is very low, especially for puppies aged 2 to 3 months. It is reported that dogs, horses, rabbits, pigs, donkeys and other animals are used to make antiviral serum, and the purified canine parvovirus enteritis immunoglobulin injection is used to treat sick dogs. Since Yokoyama e...

Claims

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Application Information

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IPC IPC(8): A61K39/395C07K16/08A61P31/14
Inventor 刘永德唐峰叶陈梁汪毅
Owner SHANGHAI JIAO TONG UNIV
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