Vaccines and immunoglobulins targeting African swine fever virus and methods of making and using same
An immunoglobulin and African swine fever technology, which is applied in the fields of vaccines and immunoglobulins targeting African swine fever virus and their preparation and use, which can solve the problems of unidentified protective antigens and lack of understanding.
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[0070] ASFV vaccine
[0071]The present disclosure generally relates to an ASFV vaccine comprising intact live ASFV particles in combination with naturally expressed ASF virus components, optionally diluted in sterile buffer, eg, sterile saline solution to about 10%. The ASFV vaccine can be used to actively immunize or vaccinate a non-susceptible species host for the production of ASFV-specific immunoglobulin. A non-susceptible species host can be a non-swine mammalian host, such as avian, equine, bovine, donkey, goat or rabbit.
[0072] Another aspect of the present disclosure relates generally to a method of producing an ASFV vaccine. In a preferred embodiment, the ASFV antigens are obtained from ASF-infected pigs or wild boars. In one embodiment, blood can be drawn from the ASF-infected pig or wild boar and collected in a blood collection tube with anticoagulant. The blood collection tube can be centrifuged, for example, at about 4°C at about 1,500 x g for about 15 minu...
Embodiment 1
[0126] On days 1, 14 and 28, three groups of chickens (n=3 per group) were immunized with the ASFV vaccine. Group 1 received saline (no vaccine) as a control, Group 2 received ASFV vaccine formulation 1 containing intact virions and immunosuppressive protein factors, and Group 3 received ASFV vaccine formulation 1 containing intact virions, viral components and immunosuppressive proteins Factor ASFV vaccine formulation 2. After the second and third immunization, blood samples were obtained and ASFV-specific antibody titers were assessed using recombinant ASFV major capsid protein p72-coated ELISA plates (SEQ ID NO: 2). The results of Example 1 demonstrate that the ASFV vaccine is immunogenic and immunization is achieved. Furthermore, Example 1 demonstrated that after 14 days ( Figure 4A ) and after 28 days ( Figure 4B ), the ASFV vaccine induced antibody pools with overall specificity for ASF viral components such as the ASFV major capsid protein p72 (SEQ ID NO: 2).
Embodiment 2
[0128] ASFV vaccine compositions were prepared from homogenates of ASFV-infected spleens from ASFV-infected pigs and ASFV-infected buffy coat containing PBMCs. The PBMC mixture was frozen in a dry ice ethanol bath and thawed to room temperature. The freeze-thaw procedure was repeated twice. The ASFV vaccine compositions were evaluated for active ASFV using qPCR. The results confirmed that the ASFV vaccine composition did not contain ASFV DNA. Three groups of laying hens (n=3 / group) were dosed with a control or one of two different ASFV vaccine formulations. Group 1 received saline as a control (no vaccine), group 2 received ASFV vaccine formulation 1 (prepared from SMNC), and group 3 received ASFV vaccine formulation 2 (prepared from SMNC and PBMC). The hens were actively immunized on days 1, 14 and 3 by administering ASFV vaccine (by intramuscular injection) or administering a control. Eggs were collected daily after the third immunization. Immunoglobulins were extracted...
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