can we generate antibody for lipid peptide conjugate, which recognize conjugate but not lipid molecule or peptide alone? i means generation of antibody specific only for lipid-peptide conjugate only.
If you immunize with the conjugate, you will get a mixture of antibodies against different epitopes. You can either make monoclonal antibodies and screen for those that recognize the desired epitope, or clone the immune response into a phage display vector to select the VL/VH pairs that recognizes the desired epitope.
thanks for answer. do you know any reference where author did such work? I mean reference for making conjugate specific antibody.
Which lipid you are taking about? I have developed polyclonal antibodies against steroids. Recent paper attached.
If any clarification is required kindly get back to me.
With best wishes.
As Annemarie Honegger has already said, you will upon immunization get a mixture of antibodies against different epitopes. What you can do is then employ a subtractive isolation protocol where you first expose your antibody pool to the lipid, then to the protein, then finally to the lipid-protein conjugate to isolate binders that only specifically recognize the conjugate.
You can also use biotinylated peptide-lipid conjugates and isolate library members with phage display. If you add in extra unconjugated lipid or peptide during later panning rounds, this is another way to remove unwanted binders (to lipid or peptide). The stringency can then be controlled by adjusting the concentration of the competitors. However, be careful to not to loose candidates at R1.
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