Logic wrote:
"No Lagat's sample is not controlled like the ones I linked to. His sample was experimental - that's the whole point of the test. If his sample was a control then there wouldn't have been any point to running the sample. A control is sample that gives a known result. Talk about obtuse. You've spoken in a court of law as an expert? Seriously?"
Any EPO test is a controlled test. There are both experimental samples and control samples run. One of the fundamental problems with Lagat's test, and ALL EPO test run prior to that, was that they did not control for degradation. It was not until Heid's scathing report that a degradation control was added. That said, if you would simply look up Lagat's test you would easily see that Lagat's test lane, and a known positive, do not appear identical at all.
If a recombinate-specific monoclonal antibody were used, a false-positive would not result due to degradation, only a false-negative would be possible. If a 2-D process rather than a 1-D process were used, degradation resulting in a charge shift would not result in a false positive unless it also resulted in a matching weight shift, so the odds of a false-positive would be greatly reduced. Yet ten years after this test was developed neither is done.
While protein stored at -20C will be stable for several years and that stored at -80C for many more, instability may result upon thawing if the sample is not properly maintained during testing. While protease inhibitors are added to the sample, these chemicals do not necessarily inhibit glycosidases, which are capable of removing the sugar groups which, to a large extent, determine the charge differences between endogenous and exogenous EPO. A standard and rational step to take to reduce the likelihood of this would be to run the samples in refrigerated centrifuges and other equipment, which would inhibit all degrading enzymes to some extent. Yet despite taking over 48 hours to run this test through all of its many steps, it is run at room temperature despite the equipment being capable of refrigeration. This is yet another factor which may result in uncertainty in the results if degrading enzymes are present.
"Talk about blathering! You sure can type a lot of words without saying anything. Until you show a controlled experiment that shows a sample that is known to have undergone degradation (not just speculation), you haven't addressed the point."
Multiple experts agreed with me, hence Lagat was cleared. How can you, a random anonymous internet poster arguing about a test they don't even bother to understand is not SDS-PAGE before running their mouth, pretend to know better?
"What does this have to do with the effect of sample degradation on the test?"
If you were educated enough on protein biochemistry to understand what I had written, the connection would be obvious. I would explain further but you have made if very clear that you are interested in arguing, not learning reality.
"I have educated myself. But no thanks to you."
Unlike other posters here, who actually looked up the test and provided useful information you could learn from, you have not brought in any factual information.
" From your first post you've been completely arrogant, rude, and obnoxious and you're contributions to the thread have been minimal except to brag about how smart you are. You must be real fun at parties."
Stating actual experience with the test and a knowledge base to comment from is not arrogance. But I do not suffer fools well, you are right for once. While it has been entertaining in a sad way, I think I'm done here, this is the reason nobody reads these boards anymore.