ChemRxiv
These are preliminary reports that have not been peer-reviewed. They should not be regarded as conclusive, guide clinical practice/health-related behavior, or be reported in news media as established information. For more information, please see our FAQs.
1/1
0/0

Assessing Accuracy of an Analytical Method in silico: Application to “Accurate Constant via Transient Incomplete Separation” (ACTIS)

preprint
revised on 05.06.2020 and posted on 08.06.2020 by Jean Luc Rukundo, J. C. Yves LeBlanc, Sven Kochmann, Sergey N. Krylov

Analytical methods may not have reference standards required for testing their accuracy. We postulate that accuracy of an analytical method can be assessed in the absence of reference standards in silico if the method is built upon deterministic processes. A deterministic process can be precisely computer-simulated thus allowing virtual experiments with virtual reference standards. Here, we apply this in silico approach to study “Accurate Constant via Transient Incomplete Separation” (ACTIS), a method for finding the equilibrium dissociation constant (Kd) of protein–small molecule complexes. ACTIS is based on a deterministic process: molecular diffusion of the interacting protein–small molecule pair in a laminar pipe flow. We used COMSOL software to construct a virtual ACTIS setup with a fluidic system mimicking that of a physical ACTIS instrument. Virtual ACTIS experiments performed with virtual samples — mixtures of a protein and a small molecule with defined rate constants and, thus, Kd of their interaction — allowed us to assess ACTIS accuracy by comparing the determined Kd value to the input Kd value. Further, the influence of multiple system parameters on ACTIS accuracy was investigated. Within multi-fold ranges of parameters, the values of Kd did not deviate from the input Kd values by more than a factor of 1.25 strongly suggesting that ACTIS is intrinsically accurate and that its accuracy is robust. Accordingly, further development of ACTIS can focus on achieving high reproducibility and precision. We foresee that in silico accuracy assessment, demonstrated here with ACTIS, will be applicable to other analytical methods built upon deterministic processes.

Funding

Natural Sciences and Engineering Research Council of Canada (NSERC) STPG-P 521331-2018

History

Email Address of Submitting Author

skochman@yorku.ca

Institution

York University, Toronto

Country

Canada

ORCID For Submitting Author

0000-0001-7423-4609

Declaration of Conflict of Interest

No conflict of interest

Version Notes

Added another model and evaluation for an extremely optimized case. Fixed some small typos and problems with embedded images in the PDF (e.g. Figure 2).

Exports