I am curious as to whether the difference in amplification signal (due to many copies of pVIII recognized by the anti-M13 antibody vs using a secondary that only recognizes the scFv would result in screening out lower affinity mAbs from the primary screen stage. Or, do you see similar hit rates with both ELISA methods?
In my experience hit rate of periplasmic ELISA tends to be lower than that of phage ELISA (sometimes much lower). I think this is not only a consequence of the difference in sensitivity due to anti-PVIII amplification, but also of the fact that some active phage-displayed scFv can never be produced as active sluble proteins in periplasm due to aggregation effects and so on.
If your final goal is to obtain bacterial scFvs, I would recommend to go directly to periplasmic ELISA to identify the best candidates.
But if you are planning to construct whole antibodies or similar molecules and express them in mammalian cells, identifying from phage ELISA could be advantageous as some molecules work pretty well in both phage display and mammalian expression, even if they are not properly folded in periplasm. Phage ELISA can help you to avoid loosing those molecules in the primary screening.
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