I need to know which is the right scale of priority when testing a mAb for possible cytotoxicity effects: which methods to use to test for the direct effect (non dependable on CDC/ADCC mechanisms) and if they are positive, then how to proceed to the next methods? Our antibodies are murine.
Hi Andrei,
In preclinical antibody screening & drug development at iQ Biosciences, we first confirm that our antibody can bind the target antigen specifically by flow cytometry to help identify both positive and negative cell lines that can be used for subsequent functional assays. We then look to examine three different types of cytotoxicity that can be mediated by your new antibody to tumor cell lines such as 1) Direct apoptotic effect of the naked antibody inducing cell death or internalization, 2) complement mediated cytotoxicity (CDC), and 3) antibody-dependent cell-mediated cytotoxicity (ADCC). These are listed with increasing technical difficulty in mind. A hint about where your antibody begin can be the isotype of your antibody as this can indicate what effector functions are more likely to be in play (IgM are better for complement; IgG1 for ADCC, etc).
For direct (non CDC / ADCC assays), I would start with incubating your cells with your antibody to see if any direct cell death can be induced. This can be measured via several color-metric dyes (CCK8, CCK-SK, etc) out that there that are optimized for adherent or suspension cells. Internalization of your antibody may also good to examine as a precursor to antibody drug conjugation (ADC). If you get stuck, iQ Biosciences offers all these screening services to help. http://www.iqbiosciences.com/bioservices/in-vitro_bioservices
Best Regards,
Omar
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