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A facile and precise method for quantifying small–large/light-weighted molecular interaction system


It is significant to quantify the intermolecular physisorption extent in biomedical field. By taking the advantage of a significant difference from either sizes or weights, we introduced a combination of Scatchard equation and either ultracentrifugation or size exclusion chromatography to obtain both the binding constant and the number of binding sites by using bovine serum albumin and eosin B as models. Compared to the photoluminescence quenching-based methods like Stern–Volmer and Hill equations, the introduced method is not only more precise but also simpler and more straightforward for the operation. Moreover, the protein conformational changes and the corresponding theoretical binding mode with an atomic resolution were also studied by using three-dimensional fluorescence spectroscopy and molecular docking method, respectively. These comparative results could help scientists select right methods to study any interactions between two molecules with significant differences from either sizes or weights.

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Corresponding author

Correspondence to Feng Zhang.

Additional information

This work was supported by the National Natural Science Foundation of China (Nos. 21171086 and 81160213), the Inner Mongolia Autonomous Region science and Technology Department (No. 211-202077), the Inner Mongolia Grassland Talent (No. 108-108038), the Natural Science Foundation of Inner Mongolia Autonomous Region of China (Nos. 2013MS1121, 2015ms0806 and 2016MS0211) and the Inner Mongolia Agricultural University (Nos. 109-108040, 211-109003 and 211-206038).

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Li, WR., Gong, P., Ma, YX. et al. A facile and precise method for quantifying small–large/light-weighted molecular interaction system. NUCL SCI TECH 27, 109 (2016).

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  • Bovine serum albumin
  • Eosin B
  • Binding constant
  • Ultracentrifugation
  • Scatchard equation
  • Size exclusion chromatography