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Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument

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Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument. / Bucciarelli, Saskia; Midtgaard, Søren Roi; Pedersen, Martin Nors; Skou, Søren; Arleth, Lise; Vestergaard, Bente.

In: Journal of Applied Crystallography, Vol. 51, No. 6, 2018, p. 1623-1632.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Bucciarelli, S, Midtgaard, SR, Pedersen, MN, Skou, S, Arleth, L & Vestergaard, B 2018, 'Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument' Journal of Applied Crystallography, vol. 51, no. 6, pp. 1623-1632. https://doi.org/10.1107/S1600576718014462

APA

Bucciarelli, S., Midtgaard, S. R., Pedersen, M. N., Skou, S., Arleth, L., & Vestergaard, B. (2018). Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument. Journal of Applied Crystallography, 51(6), 1623-1632. https://doi.org/10.1107/S1600576718014462

Vancouver

Bucciarelli S, Midtgaard SR, Pedersen MN, Skou S, Arleth L, Vestergaard B. Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument. Journal of Applied Crystallography. 2018;51(6):1623-1632. https://doi.org/10.1107/S1600576718014462

Author

Bucciarelli, Saskia ; Midtgaard, Søren Roi ; Pedersen, Martin Nors ; Skou, Søren ; Arleth, Lise ; Vestergaard, Bente. / Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument. In: Journal of Applied Crystallography. 2018 ; Vol. 51, No. 6. pp. 1623-1632.

Bibtex

@article{e75a580ebb1e47969c3835909103dc8c,
title = "Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument",
abstract = "Coupling of size-exclusion chromatography with biological solution small-angle X-ray scattering (SEC-SAXS) on dedicated synchrotron beamlines enables structural analysis of challenging samples such as labile proteins and low-affinity complexes. For this reason, the approach has gained increased popularity during the past decade. Transportation of perishable samples to synchrotrons might, however, compromise the experiments, and the limited availability of synchrotron beamtime renders iterative sample optimization tedious and lengthy. Here, the successful setup of laboratory-based SEC-SAXS is described in a proof-of-concept study. It is demonstrated that sufficient quality data can be obtained on a laboratory instrument with small sample consumption, comparable to typical synchrotron SEC-SAXS demands. UV/vis measurements directly on the SAXS exposure cell ensure accurate concentration determination, crucial for direct molecular weight determination from the scattering data. The absence of radiation damage implies that the sample can be fractionated and subjected to complementary analysis available at the home institution after SEC-SAXS. Laboratory-based SEC-SAXS opens the field for analysis of biological samples at the home institution, thus increasing productivity of biostructural research. It may further ensure that synchrotron beamtime is used primarily for the most suitable and optimized samples.",
author = "Saskia Bucciarelli and Midtgaard, {S{\o}ren Roi} and Pedersen, {Martin Nors} and S{\o}ren Skou and Lise Arleth and Bente Vestergaard",
year = "2018",
doi = "10.1107/S1600576718014462",
language = "English",
volume = "51",
pages = "1623--1632",
journal = "Journal of Applied Crystallography",
issn = "0021-8898",
publisher = "Wiley-Blackwell",
number = "6",

}

RIS

TY - JOUR

T1 - Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument

AU - Bucciarelli, Saskia

AU - Midtgaard, Søren Roi

AU - Pedersen, Martin Nors

AU - Skou, Søren

AU - Arleth, Lise

AU - Vestergaard, Bente

PY - 2018

Y1 - 2018

N2 - Coupling of size-exclusion chromatography with biological solution small-angle X-ray scattering (SEC-SAXS) on dedicated synchrotron beamlines enables structural analysis of challenging samples such as labile proteins and low-affinity complexes. For this reason, the approach has gained increased popularity during the past decade. Transportation of perishable samples to synchrotrons might, however, compromise the experiments, and the limited availability of synchrotron beamtime renders iterative sample optimization tedious and lengthy. Here, the successful setup of laboratory-based SEC-SAXS is described in a proof-of-concept study. It is demonstrated that sufficient quality data can be obtained on a laboratory instrument with small sample consumption, comparable to typical synchrotron SEC-SAXS demands. UV/vis measurements directly on the SAXS exposure cell ensure accurate concentration determination, crucial for direct molecular weight determination from the scattering data. The absence of radiation damage implies that the sample can be fractionated and subjected to complementary analysis available at the home institution after SEC-SAXS. Laboratory-based SEC-SAXS opens the field for analysis of biological samples at the home institution, thus increasing productivity of biostructural research. It may further ensure that synchrotron beamtime is used primarily for the most suitable and optimized samples.

AB - Coupling of size-exclusion chromatography with biological solution small-angle X-ray scattering (SEC-SAXS) on dedicated synchrotron beamlines enables structural analysis of challenging samples such as labile proteins and low-affinity complexes. For this reason, the approach has gained increased popularity during the past decade. Transportation of perishable samples to synchrotrons might, however, compromise the experiments, and the limited availability of synchrotron beamtime renders iterative sample optimization tedious and lengthy. Here, the successful setup of laboratory-based SEC-SAXS is described in a proof-of-concept study. It is demonstrated that sufficient quality data can be obtained on a laboratory instrument with small sample consumption, comparable to typical synchrotron SEC-SAXS demands. UV/vis measurements directly on the SAXS exposure cell ensure accurate concentration determination, crucial for direct molecular weight determination from the scattering data. The absence of radiation damage implies that the sample can be fractionated and subjected to complementary analysis available at the home institution after SEC-SAXS. Laboratory-based SEC-SAXS opens the field for analysis of biological samples at the home institution, thus increasing productivity of biostructural research. It may further ensure that synchrotron beamtime is used primarily for the most suitable and optimized samples.

U2 - 10.1107/S1600576718014462

DO - 10.1107/S1600576718014462

M3 - Journal article

VL - 51

SP - 1623

EP - 1632

JO - Journal of Applied Crystallography

JF - Journal of Applied Crystallography

SN - 0021-8898

IS - 6

ER -

ID: 208818708