On-line determination by small angle X-ray scattering of the shape of hen egg white lysozyme immediately following elution from a hydrophobic interaction chromatography column.
On-line determination by small angle X-ray scattering of the shape of hen egg white lysozyme immediately following elution from a hydrophobic interaction chromatography column.
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从疏水相互作用色谱柱洗脱后立即通过小角 X 射线散射在线测定鸡蛋清溶菌酶的形状。
DOI：
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发表时间：
2016
期刊：
影响因子：
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通讯作者：
M. Hearn
中科院分区：
文献类型：
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作者：
Chadin Kulsing;András Komáromy;R. Boysen;M. Hearn
本研究记录了使用一种集成的方法，涉及在线疏水相互作用色谱与小角X射线散射（SAM-SAXS）测量接口，以监测蛋白质的构象状态后，立即从不同温度下运行的色谱柱洗脱。此外，这种方法提供了一种额外的途径来询问可能在洗脱的色谱峰上发生的蛋白质形状的变化。为此，回转半径外推的Guinier近似与SAXS数据，而对分布函数和珠模型模拟生成的间接转换程序GNOME和从头重建与GASBOR，以提供进一步了解蛋白质的构象变化，发生在疏水相互作用色谱。
This study documents the use of an integrated approach, involving on-line hydrophobic interaction chromatography interfaced with Small Angle X-ray Scattering (HIC-SAXS) measurements, to monitor the conformational status of proteins immediately upon elution from a chromatographic column operated at different temperatures. Moreover, this approach provides an additional avenue to interrogate the changes in protein shape that may occur across the eluted chromatographic peak. To this end, radii of gyration were extrapolated from the Guinier approximation with the HIC-SAXS data, whilst pair distribution functions and bead model simulations were generated by using the indirect transform program GNOM and ab initio reconstruction with GASBOR to provide further insight into protein conformational changes that occur during hydrophobic interaction chromatography.