I have a 13 amino acids peptide and I want to inject it in mice to induce immunization. The injection will be in the tail vein. After 12 days I expect to have a peak in the production of antibodies, so in the 12th day after injection I will challenge these mice and matched controls with bacteria and perform a mortality curve. I believe this kind of immunization will be protective and prolong mice survival. How much peptide should I use to induce the immune response? One dose is enough? Do I need to inject it several times? When and how much?
If everything works well, I want to produce a monoclonal antibody against this peptide and inject it after the bacteria injections in non-immunized mice to see if this monoclonal antibody works as a new treatment for infections. How much antibody should I inject to try to treat wild-type mice with a monoclonal antibody? One dose is enough? Do I need to inject it several times? When and how much?
Hi Fabiano,
Very interesting research.
I produced many monoclonals using short peptides (8-15AA). In my own expericne, it is difficult to indue immune response using short peptides. Therefore, they are nomrally conjugated to a protein crarrier such as Ovalbumin. Conjugated peptides are then made into water in oil immulsion using Complete Freund's adjuvant for one immunisation and incomplete FA in the next three immunisations. Final boost is used before spleen collection (attached table of immunisation protocol). On day 38, you run immune response test by ELISA using tail bleed. You test the immune response against the immunogen peptide by coating ELISA plates with the same peptide. However, the peptide used in ELISA is conjugated to another carriers to make sure you are not measuring the immmune respsonse against the carrier used in immunisation.
Just final note, make sure your run some bioinformatic on your peptides to make sure it has the necessary physico-chemical characterstics (e.g. antigenic,accessible and specific).
Good luck
Abdo
Thank you very much for your response, Abdo! Do you have experience with pegylated peptides? How much conjugated peptide do you have in your final solution for the immunizations? And for monoclonal antibodies, how do you calculate the dose? best, Fabiano
Hi Fabiano,
I agree with what was said above, peptides alone will not be immunogenic. either you need to couple them to a protein such as KLH, or you can try TLR agonists such as CpGs as adjuvants. Further, you should chose another route, i.m. or s.c., i.v. will probably not work. Also, you need at least one, better two boosts, every other week. Freuds adjuvant would also work as proposed by Abdo. Good luck as well.
Hi Fabiano,
I have never tried to use pegylated peptides. I am sure you could find some literature.
As for the quantity of conjugated peptide; If its not in the table i attached. 100ul of conjugated peptide plus 100ul adjuvant for each immunisation.
I have not used monoclonals theraputically before, but seen few papers which looked into this. Article Antibodies to Conserved Surface Polysaccharides Protect Mice...
Best,
Abdo
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