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Preparation method of rabbit anti-canine parvovirus 2a type polyclonal antibodies

A canine parvovirus, polyclonal antibody technology, applied in the field of virology, can solve the problem of poor control effect of mutant strains

Inactive Publication Date: 2012-07-04
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

At present, there are few reports on the production of serum induced by immunization with canine parvovirus genotype. The production of serum is only to immunize dogs with vaccines or strains of unknown genes, and its control effect on mutant strains is not good.

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  • Preparation method of rabbit anti-canine parvovirus 2a type polyclonal antibodies
  • Preparation method of rabbit anti-canine parvovirus 2a type polyclonal antibodies
  • Preparation method of rabbit anti-canine parvovirus 2a type polyclonal antibodies

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Embodiment Construction

[0013] The present invention will be described in detail below in conjunction with specific embodiments.

[0014] 1 Materials and methods

[0015] 1.1 Experimental materials

[0016] The CPV-2a strain was purchased from the Military Veterinary Research Institute of the Academy of Military Medical Sciences, and the attenuated CPV-2 strain came from a commercial vaccine strain; F81 cat kidney cells were purchased from the Cell Bank of the Chinese Academy of Sciences; 6 adult healthy New Zealand Landrace male rabbits were selected as experimental animals. The body weight was 2-3kg, and they were fed for 1 week before the experiment, and the CPV antibody in their serum was determined to be negative.

[0017] 1.2 Reagents and instruments

[0018] DMEM culture medium is the product of Invitrogen; mycoplasma-free fetal bovine serum was purchased from Hangzhou Sijiqing Biological Co., Ltd.; glutamine was purchased from Takara Company; PEG6000 (polyethylene glycol 6000) was from Chen...

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Abstract

The invention discloses a preparation method of rabbit anti-canine parvovirus 2a type polyclonal antibodies. The method comprises the following steps of: A1) performing virus amplification; A2) concentrating antigens; A3) measuring the content of antigens; A4) adopting the immunization method; and A5) purifying serum; purifying the canine parvovirus (CPV)-2a virus strain, amplifying, concentrating the obtained virus liquid with polyethylene glycol (PEG)6000, emulsifying with Freud's adjuvant, and performing subcutaneous immunizations at ten different points, wherein basic immunization and booster immunization are totally performed four times. The prepared polyclonal antibodies have high titer, the hemagglutinin inhibition (HI) value is no less than 1:10240, and the serum neutralization (SN) value is no less than 1:51200; and compared with the CPV-2 polyclonal antibodies, the neutralizing CPV-2a antibodies have a stronger effect.

Description

technical field [0001] The invention relates to a preparation method of a rabbit anti-canine parvovirus type 2a polyclonal antibody, belonging to the technical field of virology. Background technique [0002] Canine parvovirus (canine parvovirus, CPV) is a member of the feline parvovirus subgroup of the family Parvoviridae and the genus Parvovirus. Since the discovery of CPV-2, there have been CPV-2a, CPV-2b, CPV-2c(a) and CPV-2C(b) genotypes. Currently, the canine parvoviruses prevalent in my country are CPV-2a and CPV-2b, and CPV-2a is dominant. However, the clinically used vaccine is still CPV-2, which is not effective in preventing CPV-2a, which may be a major reason for the failure of immunity to canine parvovirus disease. However, studies have found that CPV-2a vaccine cannot induce high-titer antibodies in dogs, and cannot resist the infection of CPV-2a virus. Therefore, it is of certain significance to prepare CPV-2a serum to prevent and treat domestic canine parv...

Claims

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Application Information

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IPC IPC(8): C07K16/08C07K16/06
Inventor 彭广能李思琪钟志军符华林石锦江梁璐琪舒龙马晓平
Owner SICHUAN AGRI UNIV
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