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

• Tipo: Artigo em Revista Científica Internacional
• Data: 2021

Autores

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

Revista

Título: European Biophysics Journal

Vol. 50

Páginas: 429-451

ISSN: 0175-7571

Editora: Springer Nature

Indexação

ISI Web of Knowledge - 3 Citações

Scopus - 5 Citações

Outras Informações

ID Authenticus: 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.

Idioma: Inglês

Tipo (Avaliação Docente): Científica

Nº de páginas: 23