University of Lethbridge, Dept. of Chemistry and Biochemistry, Lethbridge, Alberta, Canada.
AUC Solutions, LLC, Houston, TX, USA.
Anal Biochem. 2022 Sep 1;652:114728. doi: 10.1016/j.ab.2022.114728. Epub 2022 May 21.
Multi-wavelength analytical ultracentrifugation (MW-AUC) is a recent development made possible by new analytical ultracentrifuge optical systems. MW-AUC extends the basic hydrodynamic information content of AUC and provides access to a wide range of new applications for biopolymer characterization, and is poised to become an essential analytical tool to study macromolecular interactions. It adds an orthogonal spectral dimension to the traditional hydrodynamic characterization by exploiting unique chromophores in analyte mixtures that may or may not interact. Here we illustrate the utility of MW-AUC for experimental investigations where the benefit of the added spectral dimension provides critical information that is not accessible, and impossible to resolve with traditional AUC methods. We demonstrate the improvements in resolution and information content obtained by this technique compared to traditional single- or dual-wavelength approaches, and discuss experimental design considerations and limitations of the method. We further address the advantages and disadvantages of the two MW optical systems available today, and the differences in data analysis strategies between the two systems.
多波长分析超速离心（MW-AUC）是一种新的分析超速离心光学系统带来的最新发展。MW-AUC 扩展了 AUC 的基本流体力学信息含量，并为生物聚合物特性的广泛新应用提供了途径，有望成为研究大分子相互作用的基本分析工具。它通过利用分析物混合物中可能相互作用或不相互作用的独特发色团，为传统的流体力学特性提供了一个正交的光谱维度。在这里，我们说明了 MW-AUC 在实验研究中的应用，其中增加的光谱维度的优势提供了关键信息，这些信息是传统 AUC 方法无法获得的，也是无法解决的。我们展示了与传统的单波长或双波长方法相比，该技术在分辨率和信息量方面的改进，并讨论了该方法的实验设计注意事项和局限性。我们进一步讨论了当今两种 MW 光学系统的优缺点，以及两种系统之间数据分析策略的差异。
