Doing research project -- trying to configure dosages...

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Doing research project -- trying to configure dosages...

Post by HenryH » Sat Sep 12, 2009 2:59 pm

I'm a university student and will be doing a mandatory research project this year. What I plan to do is attach a Fas (death receptor ligand) to a mesothelin antibody by binding the 2 with the protein crosslinker Glutaraldehyde. This ligand-antibody combination will then be administered to pancreatic cancer cells that overexpress mesothelin. The purpose of the experiment is to determine whether the Fas ligand on the mesothelin antibody reacts with the death receptors for Fas after the mesothelin-ligand has bound itself to the mesothelin on the cancer cell's surface. In other words, will cancer cell apoptosis occur?

The main components of the procedure have been worked out by my mentor and I. However, we are still unsure as what dosages to use of each reagent. My mentor told me to configure based on the dosages used for similar experiments, but I haven't really found any in vitro studies on PubMed that used similar reagents with a similar experimental goal.

Does anyone know of a good database of in vitro studies I could consult? The antibody itself costs $300 for 100mcg, so I don't want to just perform the experiment based on an off-hand estimation of how much to use of each reagent (e.g., antibody, fas ligand, etc.)...

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Post by mith » Fri Oct 16, 2009 5:31 am

if you know the diffusivity for the antibody and assuming it's not significantly affected by the ligand, and you know the number of receptors you can approximate ligand receptor binding using weber's disk approximation ... sk&f=false
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