Watanabe Yasushi, Inoko Yoji
National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
J Chromatogr A. 2009 Oct 30;1216(44):7461-5. doi: 10.1016/j.chroma.2009.02.053. Epub 2009 Feb 23.
Size-exclusion chromatography with on-line synchrotron radiation solution small-angle X-ray scattering optics, absorbance and/or refractive index detectors was evaluated by protein characterizations. The radius of gyration value and zero-angle scattering intensity of protein molecules eluted from the chromatography column were estimated using this measurement system. In addition, the characterization of the conformation of the eluted proteins was demonstrated for hen egg lysozyme and bovine submaxillary mucin. The present technique will be useful for not only the determination of the radius of gyration value and molecular weight of proteins with dimensions of 1-10 nm, but also for the structural characterization of the macromolecules during the chromatography.
采用在线同步辐射溶液小角X射线散射光学装置、吸光度和/或折射率检测器的尺寸排阻色谱法通过蛋白质表征进行了评估。使用该测量系统估算了从色谱柱洗脱的蛋白质分子的回转半径值和零角度散射强度。此外，还对鸡蛋清溶菌酶和牛下颌粘蛋白洗脱蛋白的构象进行了表征。本技术不仅可用于测定尺寸为1 - 10 nm的蛋白质的回转半径值和分子量，还可用于色谱过程中大分子的结构表征。
