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A heterologous antibody against giant panda canine distemper virus and its preparation method

A canine distemper virus, giant panda technology, applied in the field of antibody and antibody preparation, can solve the problems of economic loss, allergic reaction to heterologous protein rejection, expression level, production scale and purification ability lag, etc. High and safe effect

Active Publication Date: 2021-06-01
TANGSHAN YIAN BIOLOGICAL ENG CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

From December 2014 to May 2015, 5 giant pandas in Louguantai Shaanxi Qinling Giant Panda Breeding Base died of canine distemper virus, causing heavy economic losses and adverse effects
[0004] Antibody therapy is another way to mobilize the body's immune system. Studies have shown that canine distemper antibodies have a better therapeutic effect on infected animals in the early stages of infection. Obtaining and preparing heterologous antibodies becomes inevitable
However, my country's antibody industry lags behind in terms of expression level, production scale and purification capacity. The purification level of commercially available canine distemper monoclonal antibodies and heterologous polyclonal antibodies is low. For the treatment of diseases of rare animals such as giant pandas

Method used

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  • A heterologous antibody against giant panda canine distemper virus and its preparation method
  • A heterologous antibody against giant panda canine distemper virus and its preparation method
  • A heterologous antibody against giant panda canine distemper virus and its preparation method

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1: the preparation of giant panda canine distemper inactivated vaccine

[0027] (1) Preparation and inspection of seed batches of giant panda canine distemper virus strain

[0028] The giant panda canine distemper virus strain was inoculated on a monolayer of Vero cells at 0.5%, and the virus was cultured at 37°C, and the virus supernatant was harvested when the cytopathy reached 80% to 90%, and the passage was 1 generation , and so on. Obvious cytopathic changes appeared at passage 5 (see figure 1 ). According to the above method, the giant panda canine distemper virus strain was continuously passaged in Vero cells up to 30 generations. Carry out virus content determination (see Table 1) to each generation, carry out sterility test, mycoplasma test, exogenous virus test, virulence test, specificity test and immunogenicity test to 1, 10, 15, 20, 24 generations , the test results are all qualified. It is finally determined that the highest generation of t...

Embodiment 2

[0071] Example 2: Preparation and collection of horse anti-, cat anti-, dog anti-canine distemper virus hyperimmune serum

[0072] Quarantine was carried out in accordance with relevant standards, and 4 healthy antibody-negative horses, 5 healthy antibody-negative 4-6-month-old kittens and 5 healthy antibody-negative 1-year-old Beagle dogs were selected, and they were subcutaneously passed through the back of the body respectively. The method of split-point injection was used for primary immunization and multiple booster immunizations. A total of 4 immunizations were carried out, with an interval of 14 days between each time. 0.3mg, 0.4mg, 0.5mg / cat, 0.2mg, 1.0mg, 1.5mg, 2mg, 3mg / dog. Before each immunization, venous blood was collected, the serum was separated, and the neutralizing antibody titer was tested. When the neutralizing antibody titer was above 1:10,000, the blood was collected in line with high immune serum, the serum was aseptically separated, and stored at -20°...

Embodiment 3

[0086] Example 3: Safety test of purified heterologous antibody against giant panda canine distemper virus

[0087] 1. Safety test of a single dose of immunization in dogs

[0088] Divide 15 Beagle dogs into 3 groups, 5 in each group, inject purified horse-derived antibody, cat-derived antibody and canine-derived antibody intramuscularly, 1 head / dog each time, and set another 15 Beagle dogs for three The control group was used as the control group without immunization, and was observed for 14 days. The general symptoms, body weight, body temperature and injection site of the dogs were observed 1 day before immunization and within 14 days of immunization. The results showed that after the Beagle dogs were immunized with a single dose, there were no abnormal clinical manifestations locally and systemically after the injection of the horse-derived and canine-derived antibodies, normal weight gain, no abnormal body temperature, and no significant difference from the control group...

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Abstract

The invention relates to a heterologous antibody against canine distemper virus of giant pandas, especially a horse-derived antibody, which aims to be used as a heterologous antibody for the treatment and prevention of canine distemper in giant pandas. The horse-derived antibody is derived from a healthy horse that has never suffered from any infectious disease, preferably a young horse that is about 1 year old. At the same time, the present invention relates to a method for preparing a horse-derived antibody of canine distemper virus. The method prepares an inactivated vaccine for immunization by using a virulent strain of giant panda canine distemper virus with a clear source and background. The horse immune serum was obtained from the healthy horses of the sub-immunization, and purified antibodies were obtained through further purification methods such as salting out, enzyme digestion, and chromatography, which were used for the prevention and treatment of giant panda canine distemper.

Description

【Technical field】 [0001] The invention relates to an antibody and a preparation method thereof, and further relates to a canine distemper virus heterologous antibody and a preparation method thereof. 【Background technique】 [0002] Canine distemper (Canine distemper, CD) is an acute, highly contagious infectious disease caused by canine distemper virus (Canine distemper virus, CDV), with relatively high morbidity and mortality, clinically characterized by anorexia, biphasic Fever, conjunctivitis, severe gastroenteritis, and neurological symptoms are typical features. With the expansion of the host range of canine distemper infection, since 1983, the national treasure giant panda, which is known as the "living fossil", has also been reported to be infected with canine distemper and die. In 1997, Hu Guixue and others reported that canine distemper virus infected giant pandas and caused death. From December 2014 to May 2015, five giant pandas in Louguantai Shaanxi Qinling Gia...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C07K16/06A61K39/42A61P31/14
CPCA61K2039/505A61P31/14C07K16/06C07K16/1027C07K2317/20
Inventor 赵玄多张跃武英利杨维维李丽张玉新张子佳刘静史明朱明媛王溪黄韬王晓辉谷孝玉李润梁先红
Owner TANGSHAN YIAN BIOLOGICAL ENG CO LTD
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