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Monoclonal antibody based detection of immunogens of pasteurella multocida

KrishiKosh

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Title Monoclonal antibody based detection of immunogens of pasteurella multocida
 
Creator Pankaj Kumar
 
Contributor Arvind Kumar
 
Subject Blocking ELISA, Pasteurella multocida, Mnoclonal antibody
 
Description A monoclonal antibody based search was made for immunogens of Pasteurella multocida B:2 . A
panel of five anti pasteurella mutlocida monoclonal antibodies were raised. For search of immunogens, a
monoclonal antibody blocking ELISA was developed. The anti Pasteurella multocida antibodies against
epitopes of the monoclonal antibodies were quantified in the pooled serum of vaccinated/protected cattle and
buffalo sera. Sera of 2 cattle and 3 buffaloes which showed anti-Pasteurella multocida antibody titre
>2.7log10 in a monoclonal antibody based single dilution ELISA, were pooled. The anti Pasteurella
multocida antibodies in vaccinated /protected serum blocked the binding of monoclonal antibody in the
blocking ELISA and the unblocked site became available for binding of the monoclonal antibodies when the
anti-Pasteurella multocida antibodies of the vaccinated/protected serum were diluted in dilutions of the
vaccinated/protected animal serum. Higher amount of anti-Pasteurella multocida antibodies in vaccinated/
protected serum blocked sites for the monoclonal antibody up to higher dilutions of the vaccinated/protected
serum. This way, anti-pasteurella antibodies in vaccinated/protected serum for all the five monoclonal
antibodies, were quantified.The vaccinated/protected animal serum antibody titre of monoclonal antibody
HS-1 was 1.64log10, HS-2 was 2.04 log10, HS-3 and HS-6 was 1.3 log10 and HS-4 was 2.02 log10.
The epitopes of bacterial cell proteins were detected by the monoclonal antibodies on
immunoblotting. The monoclonal antibodies dentified immunogens on proteins of 17.7kDa, 28kDa, 34 and
37KDa, 44 KDa and 55.52kDa. The HS-1 identified 17.7 kDa protein, that is a product of the ptfA gene for
type 4 fimbriae which helps in colonization of host surfaces by bacteria mediated by adhesins such as
fimbriae, the HS-2 identified the product of TonBm gene, which supplies energy for iron transport, the
HS-3 identified gene for OMP-H protein which is an envelope protein and exposed on the surface, the HS-4
identified TufA gene product, translation elongation factor A, which plays role in protein synthesis and the
HS-6 identified TolC2 outer membrane efflux channel, having important role in metabolic functions. The
monoclonal antibodies identified proteins that play role in attachment, agglutination and bacterial cell
metabolism. Antibodies interfering' attachment, causing agglutination and bacterial cell metabolic
dysfunctions are expected to play important role in protective immunity. Therefore, the epitopes identified
by the monoclonal antibodies appeared to be immunogens of Pasteurella multocida B:2.
 
Date 2016-08-17T09:22:53Z
2016-08-17T09:22:53Z
2014
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/72679
 
Language en
 
Format application/pdf
 
Publisher LUVAS