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Title: An optics-based variable-temperature assay system for characterizing thermodynamics of biomolecular reactions on solid support

Journal Article · · Review of Scientific Instruments
DOI:https://doi.org/10.1063/1.4826352· OSTI ID:22251550
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  1. Department of Chemistry, University of California, One Shields Avenue, Davis, California 95616 (United States)

A biological state is equilibrium of multiple concurrent biomolecular reactions. The relative importance of these reactions depends on physiological temperature typically between 10 °C and 50 °C. Experimentally the temperature dependence of binding reaction constants reveals thermodynamics and thus details of these biomolecular processes. We developed a variable-temperature opto-fluidic system for real-time measurement of multiple (400–10 000) biomolecular binding reactions on solid supports from 10 °C to 60 °C within ±0.1 °C. We illustrate the performance of this system with investigation of binding reactions of plant lectins (carbohydrate-binding proteins) with 24 synthetic glycans (i.e., carbohydrates). We found that the lectin-glycan reactions in general can be enthalpy-driven, entropy-driven, or both, and water molecules play critical roles in the thermodynamics of these reactions.

OSTI ID:
22251550
Journal Information:
Review of Scientific Instruments, Vol. 84, Issue 11; Other Information: (c) 2013 AIP Publishing LLC; Country of input: International Atomic Energy Agency (IAEA); ISSN 0034-6748
Country of Publication:
United States
Language:
English