trying to show that an IL6 antibody drug is effective at lowering IL6 levels/signalling in breast cancer cells. i wanna run media from cells that have received an IL6 antibody as treatment on an ELISA. the antibody should neutralize IL6 in the cell media i am testing but if i run this as a sample on the elisa then wont the ELISA just detect the antibody and give a false result. i think ive encountered this in the literature on the antibody drug siltuximab but i dont really understand the mechanism of why it wont work
It's could work if the antibodies used in ELISA didn't recognize the same epitope as the same epitope as the neutralzing antibody.
In principle you can establish two different ELISA, one showing the total IL6, if you are using a combination of antibodies as mentioned before. A second ELISA could be a competitive one, using an antibody regognizing the same epitope. This assay will give you the "available" IL6.
Just simply thry the first version in a setup adding to a constant IL6 concentration increasing concentrations of your antibody using in the treatment. Make sure that you have at least the same concentration in the ELISA setup as in your cell culture (or later therapeutic model).
It's could work if the antibodies used in ELISA didn't recognize the same epitope as the same epitope as the neutralzing antibody.
In principle you can establish two different ELISA, one showing the total IL6, if you are using a combination of antibodies as mentioned before. A second ELISA could be a competitive one, using an antibody regognizing the same epitope. This assay will give you the "available" IL6.
Just simply thry the first version in a setup adding to a constant IL6 concentration increasing concentrations of your antibody using in the treatment. Make sure that you have at least the same concentration in the ELISA setup as in your cell culture (or later therapeutic model).
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