SEC-SANS: size exclusion chromatography combined in situ with small-angle neutron scattering

Abstract

The first implementation and use of anin situsize exclusion chromatography (SEC) system on a small-angle neutron scattering instrument (SANS) is described. The possibility of deploying such a system for biological solution scattering at the Institut Laue–Langevin (ILL) has arisen from the fact that current day SANS instruments at ILL now allow datasets to be acquired using small sample volumes with exposure times that are often shorter than a minute. This capability is of particular importance for the study of unstable biological macromolecules where aggregation or denaturation issues are a major problem. The first use of SEC-SANS on ILL's instrument D22 is described for a variety of proteins including one particularly aggregation-prone system.

2017 publication guidelines for structural modelling of small-angle scattering data from biomolecules in solution: an update

“…Developments of inline purification of samples using sizeexclusion chromatography (SEC) at synchrotron SAXS beamlines (see, for example, Brennich et al, 2017;David & Pé rez, 2009;Graewert et al, 2015;Mathew et al, 2004) and at SANS beamlines (Jordan et al, 2016) is becoming increasingly popular. These SEC-SAS measurements involve the collection of SAS data as the solution elutes from the SEC column, and thus enable the separation of components of mixtures and polydisperse solutions.…”

“…Furthermore, the wwPDB Integrative/Hybrid Methods (IHM) Validation Task Force was formed (Sali et al, 2015) to address the complex issues concerning the archiving and validation of models of biomolecular complexes and assemblies that depend upon computational methods and data from independent experimental techniques, including SAS. There also have been substantial advances in analysis tools for SAS (Franke et al, 2015;Rambo & Tainer, 2013b;Schneidman-Duhovny et al, 2013;Petoukhov & Svergun, 2015;Konarev & Svergun, 2015;Petoukhov et al, 2012;Chen & Hub, 2015;Spinozzi et al, 2014;Bizien et al, 2016) and instrumentation, in particular the growth of SAS experiments utilizing inline purification and characterization (Blanchet et al, 2015;Jordan et al, 2016;Graewert et al, 2015;Brookes et al, 2013Brookes et al, , 2016Bras et al, 2014;Meisburger et al, 2016;Ibrahim et al, 2017). In regard to modelling SAS data, there has been significant increased interest and methods development in multistate/ensemblebased methods for flexible biomolecules (Tria et al, 2015;Berlin et al, 2013;Schneidman-Duhovny et al, 2016;Perkins et al, 2016;Terakawa et al, 2014) and structural modelling based on combined SAS and NMR data (Schwieters & Clore, 2014).…”

A novel experimental approach for nanostructure analysis: simultaneous small-angle X-ray and neutron scattering

“…Thus, further developments, including sample environments, multi-sample changers, optimized routine operation and userfriendly software, should be considered in future research. Some of the essential sample environments (Jordan et al, 2016;Lopez et al, 2018;Wang et al, 2004;Weigandt et al, 2011) that are currently employed at ILL could be modified/ developed to fit our system and to satisfy the demand of the ILL user community. However, a multiple-sample changer with temperature control should be constructed for routine sample measurements.…”

Biological small-angle neutron scattering: recent results and development

“…The field of biological sample environments for SANS is very dynamic and a number of innovative developments have research papers recently been presented, including online fluorescence (Ibrahim et al, 2017), SEC-SANS (Jordan et al, 2016) and microfluidics (Adamo et al, 2017). We anticipate that other laboratory biophysical techniques, such as SEC-MALLS, DLS, SLS, CD etc., or tabletop SAXS instruments will be applied to biological samples on SANS beamlines shortly.…”