Seeking to obtain Mabs to Trypsin but i want to do it with peptides. Which amino acidic sequences are more immunogenic?
This is a very difficult question. BTW: Most antibodies obtained by peptide immunization do not work with the native protein.
it`s true, i have a litle experience about it, but trypsin its very expensive for me, that´s the reason why i can`t afford it. Of course that the screening of positives clones I would do with native protein. However, thank you!!
Perhaps there is a misunderstanding about the target. Trypsin is one of the cheapest proteins (you can get it from Sigma-Aldrich or others) and therefore it should be much cheaper to use the protein than any peptide approach.
I apologize!!! but i´m a cuban scientific and we are a locked country and we can´t buy anything from American companies. For that reason it´s for us more feasible synthesize peptides for immunization and leave the little that we have trypsin to screening. I hope I expressed myself well and that you understood me
There should be companies from many countries offering trypsin. Even if you can not buy it, I would prefer to isolate it e.g from bovine pancreas. There are many protocols available. However, I did not try them myself.
Example: http://www.sciencedirect.com/science/article/pii/0003269778901732
first compare in aligment sequences from mouse an human (if you want a anti-human trypsine mAb) and look at most divergent regions between immunogene and endogenous orthologue of the "self". As several related trypsinogen sequence existe you must also consider paralogous sequences (dupmlicted sequences from the same specie) to define required specificity of the mAbs but also the "self" repertory of the host.
So "what do you want in this contexte" before "how I'm can do it".
best wish
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