Technical Notes

Validation of Tycho NT.6 precision and repeatability

Issue link:

Contents of this Issue


Page 0 of 2

TECHNICAL NOTE Validation of Tycho NT.6 precision and repeatability Introduction and Results Tycho TM NT.6 tells you so much about the quality of your protein — presence, purity, concentration, functionality and similarity — in a single experiment. It does this all by examining your protein's structural integrity or foldedness in a label-free way. The system measures the fluorescence of instrinsic tryptophan and tyrosine residues detected at both 350 nm and 330 nm as a specific defined temperature ramp is applied. Tycho generates thermal unfolding profiles and identifies the inflection temperature (T i ) that represent unfolding transition(s) or discrete changes in a protein's structural integrity. It records these unfolding profiles so you can study your protein in real-time or use the data as a reference to compare and validate the quality of your sample to any future batches. In this study we demonstrate the outstanding repeatability and intermediate precision of measurements with Tycho for both raw data and analyzed results. A reference protein, streptavidin at a concentration of 1 mg/mL, was measured on 13 different Tycho systems over a period of two weeks. Each run consisted of all six capillaries filled with 10 μL streptavidin, and for each system, 3 replicate runs were performed. In total, 234 capillaries were analyzed. We use the following definitions for repeatability and intermediate precision: Repeatability describes how similar results are when always obtained by the same operator on the same measuring system in the same lab. It is also called intra-assay precision. Beate Kern, Anna Schulze NanoTemper Technologies GmbH Floessergasse 4 81369 Munich

Articles in this issue

view archives of Technical Notes - Validation of Tycho NT.6 precision and repeatability