Preparation method of I-type duck hepatitis refined yolk antibodies

A technology for egg yolk antibody and duck hepatitis, which is applied in the direction of antiviral immunoglobulin and immunoglobulin from eggs, etc., can solve the problem of high lipid residue in egg yolk antibody, and achieve the effect of outstanding protection rate and high potency

Inactive Publication Date: 2016-03-02
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims at the technical defects of the prior art, and provides a method for preparing the refined yolk antibody against type I duck hepatitis, so as to solve the technical problem of relatively high lipid residues in the yolk antibody prepared by related methods in the prior art

Method used

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  • Preparation method of I-type duck hepatitis refined yolk antibodies
  • Preparation method of I-type duck hepatitis refined yolk antibodies
  • Preparation method of I-type duck hepatitis refined yolk antibodies

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Experimental program
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Embodiment 1

[0035] 1. Production strains

[0036] The duck hepatitis virus (Duckhepatitisvirus) type I HB-34 strain used for vaccine preparation was isolated, identified, kept and supplied by Ruipu (Baoding) Biopharmaceutical Co., Ltd.

[0037] 2. Preparation of vaccine (immunogen)

[0038] 1. Preparation of virus liquid for seedling production

[0039] Preparation of HB-34 strain virus solution Dilute the virus seed HB-34 strain 100 times, inoculate 10-day-old SPF chicken embryos into the allantoic cavity, 0.1ml per embryo, incubate at 36-37°C, and irradiate embryos twice a day examine. Chicken embryos that died within 48 to 120 hours after inoculation were placed at 2 to 8°C for 4 to 12 hours, and allantoic fluid, amniotic fluid and embryo bodies were collected, and the heads and limbs were removed from the embryo bodies. Homogenize with embryo fluid, freeze and thaw three times, centrifuge at 3000r / min for 10min, take the supernatant and mix it in a sterile container, store at 2-8°C...

Embodiment 2

[0074] 1 Immunogen preparation

[0075] 1.1 Preparation of virus liquid for seedling production

[0076] HB-34 strain venom preparation Dilute the production virus strain HB-34 100 times, inoculate 100 10-day-old SPF chicken embryos in the allantoic cavity, 0.1ml per embryo, incubate at 36-37°C, and irradiate embryos twice a day examine. Chicken embryos that died within 48 to 120 hours after inoculation were placed at 2 to 8°C for 4 to 12 hours, and allantoic fluid, amniotic fluid and embryo bodies were collected, and the heads and limbs were removed from the embryo bodies. Homogenize with embryo fluid, freeze and thaw three times, centrifuge at 3000r / min for 10min, take the supernatant and mix it in a sterile container, store at 2-8°C. (See Table 1).

[0077] Table 1 Preparation of venom

[0078] Strain name

Embryo variety

Number of embryos inoculated (pieces)

Embryo age

Harvest venom (ml)

HB-34 strain

SPF chicken embryo

100

10 ...

Embodiment 3

[0109] A kind of preparation method of type I duck hepatitis refined egg yolk antibody is characterized in that comprising the following steps:

[0110] 1) Get the egg yolk of type I duck hepatitis virus high-free eggs, stir to obtain the egg yolk liquid;

[0111] 2) Mix the egg yolk liquid and water in step 1) at a ratio of 1:0.5 (v / v), stir evenly to obtain egg yolk dilution, and keep warm at 60°C for 25 minutes;

[0112] 3) adding acetate buffer solution of 2.5 times the volume of the egg yolk liquid to the egg yolk dilution, stirring and collecting the filtrate by filter cloth;

[0113] 4) Add octanoic acid to the filtrate in step 3) to a final concentration of 3.5% (v / v), stir and place at 2°C for 4 hours;

[0114] 5) Take step 4) and filter the solution after placing until the filtrate is clarified, and collect the clarified filtrate;

[0115] 6) Take the clarified filtrate described in step 5) to filter and sterilize, then add formaldehyde to a final concentration of ...

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Abstract

The invention provides a preparation method of I-type duck hepatitis refined yolk antibodies. The preparation method comprises following steps: I-type duck hepatitis virus hyper-immune egg yolk is collected, is subjected to primary inactivation, is subjected to primary acidification extraction with an acetate buffer solution, and then is subjected to secondary inactivation taking octanoic acid as the inactivator and extraction agent; filtering clarification, filtering sterilization, and ultrafiltration concentration are carried out; and then formaldehyde is used for a third time of inactivation so as to obtain a yolk antibody solution. According to the preparation method, content of lipid in the finished product is reduced effectively via acidification water-octanoic acid method, occurrence rate of adverse reaction is reduced, the step of ultrafiltration concentration is beneficial for ensuring product antibody concentration, and increasing protection rate, in application, dilution ratio of the finished product is relatively high because of the high antibody concentration, so that impurity content is reduced indirectly, and safety is improved.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to a method for preparing a type I duck hepatitis refined egg yolk antibody. Background technique [0002] Duck viral hepatitis (Duck Virus Hepatitis, DVH) is an acute, highly lethal, severe infectious disease of ducklings caused by duck hepatitis virus (Duck Hepatitis Virus, DHV) of the family Picornaviridae Enterovirus. It is characterized by convulsions of the sick duck, head tilted back to the back, and a specific posture of opisthotonus. The main lesion is characterized by hepatomegaly, with bleeding spots and bleeding points of different sizes. Duck hepatitis virus has serotypes I, II and III, and there is no antigen correlation among the three types. In the past, the type I duck hepatitis virus was mainly prevalent in our country, but in recent years, there have been outbreaks of suspected duck hepatitis in duck populations that have been ineffective ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/10C07K16/02
Inventor 杨燚苏建东杨保收付旭彬梁武
Owner TIANJIN RINGPU BIO TECH
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