i am asking about conjugate monoclonal antibody to the surface of NH2 modified MNP i have 2 choices the first using NHS and EDC to activate the Ab first is it safe for antibody activity?
the second one is using glutaraldehyde as a crosslinker on the particles, is glutaraldehyde could react with NH2 in the Fab region ? so the antibody will loose the activity ?
which is better?
is there any precise source for the conjugation protocol?
then what about stability of functionalized particles is it required to use preservative and what is suggested preservative please
thanks :)
Dear Researcher,
The first approach would be work, I have done it previously, ECD and NHS can activate Ab and the excess reactants will be filtered after the dialysis approach.
You can visit our research as well: DOI: 10.1016/j.procbio.2021.09.006
Article Surface modified and rituximab functionalized PAMAM G4 nanop...
It is pretty unorthodox to activate an antibody with EDC/NHS or use glutaraldehyde when you are conjugating to a bead because of the risks of crosslinking the Ab. A better approach is to start with COOH-beads (these are much more common than NH2 beads) and activate with EDC/NHS. You can wash away excess EDC/NHS before adding the Ab to the activated bead.
Dr. Nick thanks for your recommendation for using glutaralhyde as a crosslinker i am afraid of binding of glutaraldhyde with NH2 terminal group in the Fab region is it affect Antibody activity ?
two step mean binding of MNP with glutaraldhyde first then adding the antibody is it right?
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