Hi All,
I am dealing with the problem of having mixed nature of monoclonal antibody in the system that is in terms of its activity of showing efficacy.
I would like to know if someone could help me to figure out, How to differentiate the nature by using any assay available.
I want clones to produce partial agonist antibody against one receptor. I have got the antibody produced as well however I am not able to figure out which kind it is. Either Full agonist, Partial agonist or Antagonistic.
Getting idea by any assay would help to choose and go for clonal expansion further and if it is not the required one then will go again with some another receptor epitope on the cell surface receptor.
Dear Santosh Kumar,
Look the links. May be useful.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3083243/
http://www.sciencedirect.com/science/article/pii/1074761395900748
Regards, Shafagat
I will start by saying that I only have limited experience in this field, but I have taken courses in which these topics were covered extensively. Broadly speaking, if you are looking to determine if the antibody is a full or partial agonist or even an antagonist, the best place to start is with assessing, in a quantitative way, the action of the native ligand with the receptor or protein of interest. Once this is done, a dose response curve can be established for the native ligand(s) with the protein of interest. Then under the same conditions you tested the functional response of protein-ligand interaction, you can then generate a dose response curve for the protein-mab interaction. If the two dose response curves get to the same level of functional output, regardless of concentration at which that maximal output is reached, then your mab can be considered a full agonist. If your mab cannot generate that same output then it is either a partial agonist or a pure antagonist. In this case, it would be best to conduct one more experiment that competes the mab with the native ligand to determine if with increasing concentrations of the mab the dose response curve either approaches that of the mab alone if it is a partial agonist or demonstrates a more purely antagonist effect. If you give a few more specifics of your experiment, I may be able to give suggestions on how to go about these experiments.
hope this helps.
Andrew
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