A multi-laboratory benchmark study of isothermal titration calorimetry (ITC) using Ca2+ and Mg2+ binding to EDTA

• Type: Article in International Scientific Journal
• Date: 2021

Authors

Velazquez Campoy, A (Author) Other; Claro, B (Author) Other; Abian, O (Author) Other; Horing, J (Author) Other; Bourlon, L (Author) Other; Claveria Gimeno, R (Author) Other; Ennifar, E (Author) Other; England, P (Author) Other; Chaires, JB (Author) Other; Wu, D (Author) Other; Piszczek, G (Author) Other; Brautigam, C (Author) Other; Tso, SC (Author) Other; Zhao, HY (Author) Other; Schuck, P (Author) Other; Keller, S (Author) Other; Bastos, M (Author) FCUP

Journal

Title: European Biophysics Journal

Vol. 50

Pages: 429-451

ISSN: 0175-7571

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 3 Citations

Scopus - 5 Citations

Other information

Authenticus ID: P-00T-S82

DOI: 10.1007/s00249-021-01523-7

Abstract (EN): A small-scale ITC benchmarking study was performed involving 9 biophysics laboratories/facilities, to evaluate inter-laboratory and intra-laboratory basal levels of uncertainty. Our prime goal was to assess a number of important factors that can influence both the data gathered by this technique and the thermodynamic parameter values derived therefrom. In its first part, the study involved 5 laboratories and 13 different instruments, working with centrally prepared samples and the same experimental protocol. The second part involved 4 additional laboratories and 6 more instruments, where the users prepared their own samples according to provided instructions and did the experiments following the same protocol as in the first part. The study design comprised: (1) selecting a minimal set of laboratories; (2) providing very stable samples; (3) providing samples not requiring preparation or manipulation; and (4) providing a well-defined and detailed experimental protocol. Thus, we were able to assess: (i) the variability due to instrument and data analysis performed by each user on centrally prepared samples; (ii) the comparability of data retrieved when using 4 different software packages to analyze the same data, besides the data analysis carried out by the different users on their own experimental results; and (iii) the variability due to local sample preparation (second part of the study). Individual values, as well as averages and standard deviations for the binding parameters for EDTA-cation interaction, were used as metrics for comparing the equilibrium association constant (logK), enthalpy of interaction (Delta H), and the so-called "stoichiometry" (n), a concentration-correction factor.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 23