Mouse polyclonals production after genetic immunization
This technique allows to make the mouse express in vivo the gene coding for the antigen of interest.
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The gene-containing plasmid is injected directly in mice that thus will express the interested antigen in vivo. Le plasmide contenant le gène d'intérêt est directement injecté chez l'animal qui va ensuite exprimer la protéine in vivo.
Among transfected cells, Antigen Presentating Cells (APC) will display the focused molecules to the immune response actors through MHC.
A part of these actors, B-cells, will produce specific antibodies.
Standard protocol :
- Construction of the DNA expression vector plasmid with a CMV promotor&enhancers containing the antigen ORF
- Transfection of HEK293 cells with plasmid DNA and preparation of antigen extract
- Injection of plasmid DNA in mice 3 times
- Bleed mice
- Test of sera for antigen specific titer using protein extract (in HEK293) as coating antigen
- If titer not satisfying, 1 or 2 other boosts
- If titer satisfying, process with rest of ascites fluid collection steps arefollowed
Key numbers :
- Minimal number of mice : …4…..
- Protocol length : 100-140…….. days
- Number of injections per animal : 3-5…..
- Quantity of antigen per injection : …20…. µg/mouse
- Number of sample collections per animal : 1 - 2…..
Related services available :
=> Gene synthesis
=> Gene cloning
=> Peptide synthesis/Recombinant production
=> Other polyclonals production
=> Monoclonals production
=> Antibody labeling
=> Antibody purification
=> Antibody fragmentation
=> Antibody refolding
=> Antibody validation