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Preparation method of riemerella anatipestifer (OmpA) monoclonal antibody cell strain

A monoclonal antibody, Riemer's duck disease technology, applied in the field of genetic engineering, can solve the problems of high mortality, economic losses in duck farming, lack of effective cross-protection of serotypes, etc., to achieve good stability and strong species. specific effect

Pending Publication Date: 2022-01-07
JIANGSU AGRI ANIMAL HUSBANDRY VOCATIONAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Riemerella anatipestifer infection, also known as duck infectious serositis, is one of the most important bacterial infectious diseases infecting poultry. The disease is susceptible to ducks aged one to eight weeks. Infected ducks will appear depressed, green Diarrhea, serous secretions from the eyes and nose, and some neurological symptoms, characterized by fibrinous pericarditis, perihepatitis, air sac inflammation and caseous salpingitis, is an infectious sepsis with high mortality, and is of great concern to the duck industry. cause large economic losses
[0003] In addition to infecting ducks, R. anatipestifer can also infect chickens, turkeys, geese and other poultry. There are many serotypes of R. anatipestifer. Twenty-one serotypes have been reported worldwide, of which serotype Types 1, 2, 6 and 10 are the main domestic serotypes, and there is no effective cross-protection between the serotypes

Method used

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  • Preparation method of riemerella anatipestifer (OmpA) monoclonal antibody cell strain

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Embodiment 1

[0027] see figure 1 , the present invention provides a technical scheme: when the preparation method of the Riemerella anatipestifer monoclonal antibody cell strain is used, the method comprises the following steps:

[0028] Step S1: preparatory procedures before preparation, preparation of bacterial strains and bacterial strains; preparation of experimental animals; preparation of reagents and instruments.

[0029] Step S2: Animal immunization, drug preparation; multi-point injection; re-injection; serum separation; detection of serum titer, the detection of serum titer is to use indirect ELISA method to detect serum titer, select the mouse with the highest titer for the second step Four times of booster immunization, the dose of immunogen is twice that of the initial immunization without using Freund's adjuvant adjuvant, and the cell fusion is carried out within 72 hours.

[0030] Step S3: cell fusion, take the mice 3 days after immunization, remove the eyeballs of the mice...

Embodiment 2

[0044] Embodiment 2, on the basis of the foregoing embodiments, another embodiment is specifically:

[0045] Further, the re-injection in step S2 is to use the purified OmpA and incomplete Freund's adjuvant to mix and emulsify the second and third immunization injections of the mice.

Embodiment 3

[0046] Embodiment 3, on the basis of the foregoing embodiments, another embodiment is specifically:

[0047] Further, the isolated serum in step S2 is two weeks after the third booster immunization, the blood of the retro-orbital vein of the mouse is collected and the serum is isolated.

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Abstract

The invention discloses a preparation method of a riemerella anatipestifer (OmpA) monoclonal antibody cell strain, and relates to the technical field of genetic engineering. The preparation method comprises the following steps of: S1, preparation before preparation: preparing a strain and a fine strain, preparing a test animal, and preparing a reagent and an instrument; S2, animal immunization: preparing a medicament, performing multi-point injection, performing secondary injection, separating serum, and detecting the titer of the serum; S3, cell fusion; S4, cell preparation; S5, completion; S6, subsequent detection; and S7, summarization. According to the preparation method, OmpA is a structural protein with high conservative property and has very high immunogenicity and immunoreactivity by using an OmpA structural protein; a hybridoma cell strain capable of stably secreting an OmpA monoclonal antibody is successfully prepared by a hybridoma technology, and the serum has an effective cross protection effect.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a method for preparing a Riemerella anatipestifer monoclonal antibody cell strain. Background technique [0002] Riemerella anatipestifer is a Gram-negative, aerobic, brevibacterium. Riemerella anatipestifer infection, also known as duck infectious serositis, is one of the most important bacterial infectious diseases infecting poultry. The disease is susceptible to ducks aged one to eight weeks. Infected ducks will appear depressed, green Diarrhea, serous secretions from the eyes and nose, and some neurological symptoms, characterized by fibrinous pericarditis, perihepatitis, air sac inflammation and caseous salpingitis, is an infectious sepsis with high mortality, and is of great concern to the duck industry. cause large economic losses. [0003] In addition to infecting ducks, R. anatipestifer can also infect chickens, turkeys, geese and other poultry. There are m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/10C12R1/91
CPCC07K16/1203C07K2317/94C07K2317/33
Inventor 朱善元于圣青董洪燕吴植封琦
Owner JIANGSU AGRI ANIMAL HUSBANDRY VOCATIONAL COLLEGE
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