Trivalent virus antibodies and preparation method thereof

A virus and antibody technology, applied in antiviral immunoglobulin, egg-derived immunoglobulin, etc., can solve the problem that the protection effect of monotype antibody cannot be guaranteed, the activity of yolk antibody is affected, and the safety of antibody is affected, so as to achieve virus elimination. Good living effect, stable biological activity, long time effect

Inactive Publication Date: 2015-08-05
CHENGDU ANTIK BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are bivalent HFMD virus egg yolk antibodies (CoxA16, EV71), but they cannot produce blocking and preventive protection against the CoxA10 HFMD virus that is prevalent in some areas, and the antigen used to prepare the antibody is a compound immune antigen (Immune the hen after mixing the bivalent antigen), the titer of the monotype antibody produced by it varies with the difference of the hen body, and the protective effect of the monotype antibody cannot be guaranteed
[0006] There are existing methods for preparing egg yolk antibodies. Although the immunized chickens are healthy laying hens that have been selected, inspected and vaccinated against certain diseases, there are still some chicken-derived bacteria or / and viruses mixed in the eggs to enter Possibility in yolk antibody, such as Escherichia coli, Salmonella, chicken egg drop syndrome virus, chicken avian leukemia virus, etc., thus affecting the safety of the antibody
At present, some studies use toxic formaldehyde, divinylamine (BEI) and β-propiolactone (BPL) as inactivators, which have inactivation effects on exogenous viruses in egg yolk antibodies, but have a negative effect on the activity of egg yolk antibodies. affect the effect of the effect

Method used

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  • Trivalent virus antibodies and preparation method thereof

Examples

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Effect test

Embodiment

[0041] The trivalent virus antibody in this example is a trivalent chicken yolk antibody against HFMD virus, and the trivalent HFMD virus includes EV71, CoxA10, and CoxA16.

[0042] The specific preparation method of the trivalent virus antibody, such as figure 1 shown, including the following steps:

[0043](1) Inoculate the EV71, CoxA10 and CoxA16 hand-foot-mouth virus strains into Vero cells, and place the Vero cells inoculated with the virus strains in a 33-36°C incubator for 5-7 days. Harvest the virus liquid, determine the specific value of CCID50 according to the received virus liquid, use 0.2-0.4% formaldehyde for inactivation, and put it in a temperature environment of 2-8°C after centrifugation and purification; according to the results of CCID50, EV71, CoxA10 and CoxA16 were diluted with buffer solution, respectively, and the same amount of Freund's complete adjuvant or Freund's incomplete adjuvant to prepare emulsified immune antigens.

[0044] (2) Take a healthy...

Embodiment 2

[0060] In this example, on the basis of the above examples, ELISA method was used to detect the titers of three monotype antibody stock solutions, and the antibody titers were measured: EV71≥1:32, CoxA16≥1:32, CoxA10≥1:32. Other parts of this embodiment are the same as those of the foregoing embodiment, and will not be repeated here.

Embodiment 3

[0062] In this example, on the basis of the above examples, in order to verify the effectiveness of the trivalent virus antibody, a virus inactivation experiment was carried out on the obtained anti-trivalent hand, foot and mouth disease virus chicken egg yolk antibody. The specific process is as follows:

[0063] with a titer of 10 5 ~10 7 The hand, foot and mouth disease virus EV71, CoxA16, CoxA10 type suspensions were diluted with 3% bovine serum albumin respectively, and 0.2ml of the diluted hand, foot and mouth disease virus EV71, CoxA16, CoxA10 type suspensions were added to sterile test tubes Add 0.8ml of chicken egg yolk antibody obtained against trivalent hand, foot and mouth disease virus, mix well, and then place it in a water bath at 20°C±1°C for 5min, 10min and 20min. After shaking and mixing, samples were taken to detect the titer of residual HFMD virus. The average inactivation log value of HFMD virus EV71, CoxA16 and CoxA10 is not less than 1.0log.

[0064] ...

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Abstract

The invention discloses trivalent virus antibodies and a preparation method thereof. The trivalent virus antibodies are trivalent chicken egg-yolk antibodies resisting hand-foot-and-mouth disease viruses, wherein the trivalent hand-foot-and-mouth disease viruses include EV71, CoxA10 and CoxA16. The preparation method comprises the following steps: (1) respectively preparing virus immunizing antigens; (2) injecting the three different virus immunizing antigens into different hens; (3) collecting immunized eggs produced by the immunized hens and extracting and purifying the egg yolks of the immunized eggs so as to respectively obtain three monotype antibody stock solutions; and (4) adding a stabilizer to the three obtained monotype antibody stock solutions according to the antibody titer determination so as to obtain the chicken egg-yolk antibodies. The trivalent virus antibodies provided by the invention are capable of neutralizing tri-type virus antigen and have wider protection coverage; monotype antibodies in a prepared tri-type antibody stock solution are capable of inhibiting and killing antigens independently; the trivalent virus antibodies are stable in bioactivity, high in titer and good in safety, and can be stored for a long time.

Description

technical field [0001] The invention relates to the technical field of biological antibody immunization, in particular to a trivalent virus antibody and a preparation method thereof. Background technique [0002] Hand, foot and mouth disease is an infectious disease in children. There are a variety of enteroviruses that can cause hand, foot and mouth disease. The most common types are CoxA16, EV71 and CoxA10. Its infection routes include digestive tract, respiratory tract and contact transmission. It mostly occurs in children under 5 years old, and the incubation period is mostly 2 to 10 days. It can cause herpes on the hands, feet, mouth and other parts. A small number of children can cause complications such as myocarditis, pulmonary edema, and aseptic meningoencephalitis. The main mode of transmission is through air droplets. If the disease develops rapidly in severely ill children, it will lead to death. For hand, foot and mouth disease, there is no preventive vaccine ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C07K16/02
Inventor 林小军叶琳朱玲李燕何丽珊万强何珊叶向阳蔡仲林烨暄
Owner CHENGDU ANTIK BIOTECH CO LTD
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