中等中子通量仪器的排粒径色谱-小角中子散射系统优化。

IF 2.8 3区 材料科学 Q1 Biochemistry, Genetics and Molecular Biology
Journal of Applied Crystallography Pub Date : 2025-02-11 eCollection Date: 2025-04-01 DOI:10.1107/S1600576725000779
Ken Morishima, Rintaro Inoue, Tatsuo Nakagawa, Masahiro Shimizu, Ritsuki Sakamoto, Tatsuro Oda, Koichi Mayumi, Masaaki Sugiyama
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引用次数: 0

摘要

除了目标生物大分子外,生物大分子溶液不可避免地含有杂质。这一直是实现高精度溶液散射测量的主要障碍。为了克服这一问题,开发了小角度x射线散射(SAXS) -粒径排除色谱(SEC-SAXS)。该方法包括将SEC洗脱的溶液直接注射到测量细胞中,并在目标生物大分子洗脱过程中进行SAXS测量。这项技术导致了生物大分子溶液散射的范式转变。目前,SEC-SAXS系统在小角中子散射(SANS)中的应用正在推进。然而,由于样品溶液中的目标生物大分子不仅被纯化,而且被SEC稀释并在短时间内通过样品细胞,因此该方法正在高中子通量的SANS仪器中实施。在此,我们开发了一种新型的SEC-SANS系统,它可以在中等中子通量的SANS仪器上有效地工作。它的关键创新是设计和优化了专用的流动路径,允许在样品细胞中仅存储从SEC洗脱的目标生物分子。这一创新实现了SEC样本的长时间测量,称为“停止模式”。因此,该方法可以获得目标生物大分子的高精度溶液散射数据,甚至可以在中等中子通量下使用SANS仪器进行SEC-SANS测量。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Size-exclusion chromatography-small-angle neutron scattering system optimized for an instrument with medium neutron flux.

Biomacromolecular solutions inevitably contain impurities in addition to the target biomacromolecules. This has been a major obstacle to achieving high-precision solution scattering measurements. To overcome this problem, small-angle X-ray scattering (SAXS) coupled with size-exclusion chromatography (SEC-SAXS) has been developed. This method involves injecting the solution eluted by SEC directly into a measurement cell and conducting SAXS measurements during the elution of the target biomacromolecule. This technique has resulted in a paradigm shift in biomacromolecule solution scattering. Currently, the application of the SEC-SAXS system to small-angle neutron scattering (SANS) is being advanced. However, since the target biomacro-mol-ecules in the sample solution are not only purified but also diluted by SEC and pass through the sample cell in a short time, this method is being implemented in SANS instruments at high neutron flux. Here, we developed a new type of SEC-SANS system that can operate effectively with a SANS instrument at medium neutron flux. Its key innovation is the design and optimization of a dedicated flow path that allows for the storage of only the target biomolecules eluted from SEC in the sample cell. This innovation enables long-duration measurements, termed the 'stopping mode', for SEC samples. Consequently, this method allows for acquiring high-precision solution scattering data for target biomacromol-ecules, enabling SEC-SANS measurements even with SANS instruments at medium neutron flux.

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来源期刊
CiteScore
10.00
自引率
3.30%
发文量
178
审稿时长
4.7 months
期刊介绍: Many research topics in condensed matter research, materials science and the life sciences make use of crystallographic methods to study crystalline and non-crystalline matter with neutrons, X-rays and electrons. Articles published in the Journal of Applied Crystallography focus on these methods and their use in identifying structural and diffusion-controlled phase transformations, structure-property relationships, structural changes of defects, interfaces and surfaces, etc. Developments of instrumentation and crystallographic apparatus, theory and interpretation, numerical analysis and other related subjects are also covered. The journal is the primary place where crystallographic computer program information is published.
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