There's an interaction with a protein, FAP2, on a bacteria, F. Nucleatum, with TIGIT on natural killer cells. I want to target the natural killer cells specifically to block this interaction between the bacteria and the NK cells. I wanted to insert an antibody in mice transgenic for human TIGIT, however if I do so this will elicit an auto-immune response. How can I get around this? I'll consider alternative approaches outside of targeting FAP2 and/or the bacteria.
You can make F(ab')2 fragments of your antibodies - there are lots of protocols for this available, but the most common way to do it is add pepsin for a few hours to cleave the Fc portion of the antibody. This will still block the receptor, but without the Fc portion it won't activate complement or Fc-receptor-mediated responses. This assumes that your antibody is a blocking antibody, and if you are not sure of this then you should test it in vitro first.
You can either make F(ab')2 fragments using the specific FabRICATOR (IdeS) protease or you could inactivate the Fc by removing the Fc-glycan using GlycINATOR (EndoS2). In the latter approach you still have binding to FcRn but not to the activating FcRs.
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