Hello Francesca,

if you are referring to immunogenicity as immunogenic for T-cells, then the panel of differentially expressed tumor antigens includes many of the ones you mentioned (you find a listing of peptide antigens in the peptide database of the cancerimmunity.org site). Before you invest a lot of money in your antibodies, I would recommend to analyze your cells on the mRNA level (which would be a lot cheaper). It is generally accepted, that T-cells potentially recognize a cell which is PCR positive, because in any case, it is not the whole protein, but the peptides derived from its degradation that are presented.

A more precise way would be to analyze your lysate for these peptides (by HPLC), but unless your are a specialist in that field, it may be quite difficult given the crude mixture such a lysate is.

Finally, if you have a T-cell clone for standard melanosomal antigens such as Melan A or Meloe or PRAME, you could use lysate from PCR+ tumor cells, pulse it onto DC and ask, whether your T-cells are activated in any way.

(and if you still want to use antibodies, think about Western Blotting rather than flow...)

Hope this helps,

Matthias

Yes, I was referring to T-cell response. Thank you, Matthias, good suggestions. I will check the database you mentioned. I have got a T cell clone positive to melan A and I will surely use it in a elispot assay to test activation by my lysates loaded on T2 cell line. I ought to buy primers...purified antiboies are not so expensive, and, also, I am a fanatic cytoflower. No way to use HPLC. :) Thank you again.

T2 cells are deficient in antigen-processing. So unless your lysate contains the exact peptide epitope which can be loaded onto the MHC complex, the immunogenic peptide is not going to be presented by T2-cells, even if DC may be able to present the antigen after additional processing.

Good luck,

Matthias