Online Ion-exchange Chromatography for Small-angle X-ray Scattering

Overview

Journal Acta Crystallogr D Struct Biol

Specialty Biochemistry

Date 2016 Oct 7

PMID 27710930

Citations 9

Authors

Stephanie Hutin

Martha Brennich

Benoit Maillot

Adam Round

Affiliations

Soon will be listed here.

Abstract

Biological small-angle X-ray scattering (BioSAXS) is a powerful technique to determine the solution structure, particle size, shape and surface-to-volume ratio of macromolecules. However, a drawback is that the sample needs to be monodisperse. To ensure this, size-exclusion chromatography (SEC) has been implemented on many BioSAXS beamlines. Here, the integration of ion-exchange chromatography (IEC) using both continuous linear and step gradients on a beamline is described. Background subtraction for continuous gradients by shifting a reference measurement and two different approaches for step gradients, which are based on interpolating between two background measurements, are discussed. The results presented here serve as a proof of principle for online IEC and subsequent data treatment.

Citing Articles

Adaptable SEC-SAXS data collection for higher quality structure analysis in solution.

Matsui T, Rajkovic I, Mooers B, Liu P, Weiss T Protein Sci. 2024; 33(4):e4946.

PMID: 38501481 PMC: 10949327. DOI: 10.1002/pro.4946.

: a general method to deconvolve X-ray scattering data from evolving mixtures.

Meisburger S, Xu D, Ando N IUCrJ. 2021; 8(Pt 2):225-237.

PMID: 33708400 PMC: 7924237. DOI: 10.1107/S2052252521000555.

Solution scattering at the Life Science X-ray Scattering (LiX) beamline.

Yang L, Antonelli S, Chodankar S, Byrnes J, Lazo E, Qian K J Synchrotron Radiat. 2020; 27(Pt 3):804-812.

PMID: 32381785 PMC: 7206542. DOI: 10.1107/S1600577520002362.

Integrated beamline control and data acquisition for small-angle X-ray scattering at the P12 BioSAXS beamline at PETRAIII storage ring DESY.

Hajizadeh N, Franke D, Svergun D J Synchrotron Radiat. 2018; 25(Pt 3):906-914.

PMID: 29714204 PMC: 5929361. DOI: 10.1107/S1600577518005398.

An endogenous dAMP ligand in class Ib RNR promotes assembly of a noncanonical dimer for regulation by dATP.

Parker M, Maggiolo A, Thomas W, Kim A, Meisburger S, Ando N Proc Natl Acad Sci U S A. 2018; 115(20):E4594-E4603.

PMID: 29712847 PMC: 5960316. DOI: 10.1073/pnas.1800356115.

References

Kikhney A, Svergun D . A practical guide to small angle X-ray scattering (SAXS) of flexible and intrinsically disordered proteins. FEBS Lett. 2015; 589(19 Pt A):2570-7. DOI: 10.1016/j.febslet.2015.08.027. View

Brookes E, Perez J, Cardinali B, Profumo A, Vachette P, Rocco M . Fibrinogen species as resolved by HPLC-SAXS data processing within the () enhanced SAS module. J Appl Crystallogr. 2013; 46(Pt 6):1823-1833. PMC: 3831300. DOI: 10.1107/S0021889813027751. View

Franke D, Jeffries C, Svergun D . Correlation Map, a goodness-of-fit test for one-dimensional X-ray scattering spectra. Nat Methods. 2015; 12(5):419-22. DOI: 10.1038/nmeth.3358. View

Rambo R, Tainer J . Accurate assessment of mass, models and resolution by small-angle scattering. Nature. 2013; 496(7446):477-81. PMC: 3714217. DOI: 10.1038/nature12070. View

Round A, Felisaz F, Fodinger L, Gobbo A, Huet J, Villard C . BioSAXS Sample Changer: a robotic sample changer for rapid and reliable high-throughput X-ray solution scattering experiments. Acta Crystallogr D Biol Crystallogr. 2015; 71(Pt 1):67-75. PMC: 4304687. DOI: 10.1107/S1399004714026959. View

De Maria Antolinos A, Pernot P, Brennich M, Kieffer J, Bowler M, Delageniere S . ISPyB for BioSAXS, the gateway to user autonomy in solution scattering experiments. Acta Crystallogr D Biol Crystallogr. 2015; 71(Pt 1):76-85. PMC: 4304688. DOI: 10.1107/S1399004714019609. View

Skou S, Gillilan R, Ando N . Synchrotron-based small-angle X-ray scattering of proteins in solution. Nat Protoc. 2014; 9(7):1727-39. PMC: 4472361. DOI: 10.1038/nprot.2014.116. View

Jensen M, Norgaard Toft K, David G, Havelund S, Perez J, Vestergaard B . Time-resolved SAXS measurements facilitated by online HPLC buffer exchange. J Synchrotron Radiat. 2010; 17(6):769-73. DOI: 10.1107/S0909049510030372. View

Selkirk C . Ion-exchange chromatography. Methods Mol Biol. 2004; 244:125-31. DOI: 10.1385/1-59259-655-x:125. View

10.

Graewert M, Svergun D . Impact and progress in small and wide angle X-ray scattering (SAXS and WAXS). Curr Opin Struct Biol. 2013; 23(5):748-54. DOI: 10.1016/j.sbi.2013.06.007. View

11.

Knight C, Hub J . WAXSiS: a web server for the calculation of SAXS/WAXS curves based on explicit-solvent molecular dynamics. Nucleic Acids Res. 2015; 43(W1):W225-30. PMC: 4489308. DOI: 10.1093/nar/gkv309. View

12.

Jeffries C, Graewert M, Svergun D, Blanchet C . Limiting radiation damage for high-brilliance biological solution scattering: practical experience at the EMBL P12 beamline PETRAIII. J Synchrotron Radiat. 2015; 22(2):273-9. DOI: 10.1107/S1600577515000375. View

13.

Mathew E, Mirza A, Menhart N . Liquid-chromatography-coupled SAXS for accurate sizing of aggregating proteins. J Synchrotron Radiat. 2004; 11(Pt 4):314-8. DOI: 10.1107/S0909049504014086. View

14.

Majorek K, Porebski P, Dayal A, Zimmerman M, Jablonska K, Stewart A . Structural and immunologic characterization of bovine, horse, and rabbit serum albumins. Mol Immunol. 2012; 52(3-4):174-82. PMC: 3401331. DOI: 10.1016/j.molimm.2012.05.011. View

15.

Graewert M, Franke D, Jeffries C, Blanchet C, Ruskule D, Kuhle K . Automated pipeline for purification, biophysical and x-ray analysis of biomacromolecular solutions. Sci Rep. 2015; 5:10734. PMC: 5377070. DOI: 10.1038/srep10734. View

16.

Watanabe Y, Inoko Y . Size-exclusion chromatography combined with small-angle X-ray scattering optics. J Chromatogr A. 2009; 1216(44):7461-5. DOI: 10.1016/j.chroma.2009.02.053. View

17.

Yigzaw Y, Hinckley P, Hewig A, Vedantham G . Ion exchange chromatography of proteins and clearance of aggregates. Curr Pharm Biotechnol. 2009; 10(4):421-6. DOI: 10.2174/138920109788488842. View

18.

Ashiotis G, Deschildre A, Nawaz Z, Wright J, Karkoulis D, Picca F . The fast azimuthal integration Python library: . J Appl Crystallogr. 2015; 48(Pt 2):510-519. PMC: 4379438. DOI: 10.1107/S1600576715004306. View

19.

Grant T, Luft J, Wolfley J, Tsuruta H, Martel A, Montelione G . Small angle X-ray scattering as a complementary tool for high-throughput structural studies. Biopolymers. 2011; 95(8):517-30. PMC: 3124082. DOI: 10.1002/bip.21630. View

20.

Pernot P, Round A, Barrett R, De Maria Antolinos A, Gobbo A, Gordon E . Upgraded ESRF BM29 beamline for SAXS on macromolecules in solution. J Synchrotron Radiat. 2013; 20(Pt 4):660-4. PMC: 3943554. DOI: 10.1107/S0909049513010431. View