A Rigorous Experiment to Determine If a Quantitative Urine Assay Suffers from Non-Specific Binding

摘要

1. A rigorous NSB experiment is imperative for a true indication of NSB. Critical elements such as using pipettes for transferring from tube to tube and evaluating multiple urine lots should be incorporated into a NSB experiment. 2. The larger the surface area, the more NSB loss; therefore, keep sample volumes ＜20% of the total tube volume for the experiment. In addition, the experiment should be performed at a concentration near the LLOQ so that NSB loss can definitively be detected. 3. The length of time does not seem to be a key factor in NSB loss; therefore, a wait step in between each tube transfer is not important. The wait step will be removed from our current approach. 4. At least six transfers should be evaluated, as only two or four transfers may not show a significant loss due to NSB. 5. The number of transfers and the significance of the observed NSB loss is partially dependent on how many transfers the study sample will experience from collection through analysis.