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Heterologous antibody of panda canine distemper virus and preparation method thereof

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

Active Publication Date: 2020-04-03
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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  • Heterologous antibody of panda canine distemper virus and preparation method thereof
  • Heterologous antibody of panda canine distemper virus and preparation method thereof
  • Heterologous antibody of panda canine distemper virus and preparation method thereof

Examples

Experimental program
Comparison scheme
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 a single dose of immunization, the horse-derived and canine-derived antibodies had no abnormal clinical manifestations locally and throughout the body, normal weight gain, and no abnormal body temperature, which was not significantly different from that of the control group of Beagle dogs. On th...

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Abstract

The invention relates to a heterologous antibody for panda canine distemper virus, in particular to a horse-derived antibody, and aims to be used as the heterologous antibody for treating and preventing canine distemper of pandas. The horse-derived antibody is derived from healthy horses which are not affected by any infectious diseases, preferably young horses which are about 1 year old. Meanwhile, the invention relates to a preparation method of the horse-derived antibody of the canine distemper virus. According to the method, a panda canine distemper virus virulent strain with clear sourceand background is used for preparing an inactivated vaccine for immunization, the inactivated vaccine is used for immunizing healthy horses for multiple times to obtain immune serum of horses, and further purification methods such as salting-out, enzyme digestion and chromatography are carried out to obtain a purified antibody which is used for preventing and treating 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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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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