Comparison of methods for quantitative biomolecular interaction analysis

Author:

Conrad Monika,Fechner Peter,Proll Günther,Gauglitz Günter

Abstract

AbstractIn order to perform good kinetic experiments, not only the experimental conditions have to be optimized, but the evaluation procedure as well. The focus of this work is the in-depth comparison of different approaches and algorithms to determine kinetic rate constants for biomolecular interaction analysis (BIA). The different algorithms are applied not only to flawless simulated data, but also to real-world measurements. We compare five mathematical approaches for the evaluation of binding curves following pseudo-first-order kinetics with different noise levels. In addition, reflectometric interference spectroscopy (RIfS) measurements of two antibodies are evaluated to determine their binding kinetics. The advantages and disadvantages of the individual approach will be investigated and discussed in detail. In summary, we will raise awareness on how to evaluate and judge results from BIA by using different approaches rather than having to rely on “black box” closed (commercial) software packages.

Funder

Eberhard Karls Universität Tübingen

Publisher

Springer Science and Business Media LLC

Subject

Biochemistry,Analytical Chemistry

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