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
