There are varying views on the choice for capture and detection Ab for immunoassay application. While a large no of people (mostly academic researchers) prefer polyclonal for capture, there is a significant no of people (industry experts) who says monoclonal should be used for capture. Similar arguments exist for detection antibody selection.
For capture Ab:
Monoclonal- reduces cross reactivity and more specific, but slow binding kinetics
Polyclonal- binds far more protein, and gives larger signal than monoclonal even after incorporating variations due to cross-reactivity
Detection/primary Ab: arguments are on similar line
Ours is a microfluidic application where binding will take place in reaction-limited regime => much less binding than conventional assay.
In this case, you should use the monoclonal antibody for capture. Otherwise, the limited surface for immobilization would not lead to a sufficient binding capacity based on the polyclonal antibody if not highly purified. However, this rule only applies, if the affinities of the Mab and the Pab are similar. For secondary (labelled) antibodies, also polyclonal ones are preferable (cost, flexibility and robustness).
Thank you once again Dr. Weller. Your suggestions are well appreciated.
Any assay that you develop for being used commercially, i.e. often and over a long time, would greatly benefit from monoclonal or even better recombinant antibodies. Also, you may be aware of recent crisis wirh polyclonal antibodies in the US which may result in shortage of supply. An often overlooked aspect is the regeneration performance required for many biosensor applications (in contrast to e.g. ELISA). Getting a different batch of the polyclonal can quickly result in problems. Finally, monoclonals / recombinants are better defined reagents that can be used to obtain more information from the biosensor experiments, hence offering additional benefits over ELISA and other endpoint assays.
I agree with Marcus. If you have the budget, time and expertise to develop monoclonal or recombinant antibodies of high quality, they are better on the long term. Regarding the comment on the "antibody crisis" and varying quality of batches, you may read this paper.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4803150/
Thank you Dr. Sack and Dr. Weller, for your inputs. I appreciate your time and help.
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