Automated microfluidic sample-preparation platform for high-throughput structural investigation of proteins by small-angle X-ray scattering

Lafleur, Josiane; Snakenborg, Detlef; Nielsen, Stefan Kragh; Møller, Magda; Toft, Katrine Nørgaard; Menzel, Andreas; Jacobsen, Jes K.; Vestergaard, Bente; Arleth, Lise; Kutter, Jörg Peter

doi:10.1107/s0021889811030068

Cited by 33 publications

(35 citation statements)

References 31 publications

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“…The UV absorption was monitored using the UV cell on the exposure chip (see Fig. 1b) following the same basic principles as given by Lafleur et al (2011). Optical fibers with a 250 mm diameter were inserted into the 250 Â 250 mm side channels on the exposure chip, giving a path length of 1 mm.…”

Section: Uv Measurementsmentioning

confidence: 99%

“…Thus, it is a requisite to analyze the structural state of a given protein under a large number of experimental conditions (Toft et al, 2008;Lafleur et al, 2011). Conventional biological SAXS experiments also require exposure at multiple protein concentrations.…”

Section: Introductionmentioning

confidence: 99%

“…An automated lab-on-a-chip microfluidic sample preparation system for SAXS has recently been presented (Toft et al, 2008;Nielsen, 2009;Lafleur et al, 2011). This chip enabled automated mixing of samples and featured a sample X-ray chamber with fiber optics for integrated ultraviolet (UV) measurements.…”

Section: Introductionmentioning

confidence: 99%

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In situ microfluidic dialysis for biological small-angle X-ray scattering

Skou

Jensen

et al. 2014

J Appl Cryst

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Owing to the demand for low sample consumption and automated sample changing capabilities at synchrotron small-angle X-ray (solution) scattering (SAXS) beamlines, X-ray microfluidics is receiving continuously increasing attention. Here, a remote-controlled microfluidic device is presented for simultaneous SAXS and ultraviolet absorption measurements during protein dialysis, integrated directly on a SAXS beamline. Microfluidic dialysis can be used for monitoring structural changes in response to buffer exchange or, as demonstrated, protein concentration. By collecting X-ray data during the concentration procedure, the risk of inducing protein aggregation due to excessive concentration and storage is eliminated, resulting in reduced sample consumption and improved data quality. The proof of concept demonstrates the effect of halted or continuous flow in the microfluidic device. No sample aggregation was induced by the concentration process at the levels achieved in these experiments. Simulations of fluid dynamics and transport properties within the device strongly suggest that aggregates, and possibly even higher-order oligomers, are preferentially retained by the device, resulting in incidental sample purification. Hence, this versatile microfluidic device enables investigation of experimentally induced structural changes under dynamically controllable sample conditions.

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Section: Uv Measurementsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In situ microfluidic dialysis for biological small-angle X-ray scattering

Skou

Jensen

et al. 2014

J Appl Cryst

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“…pH or salt concentration. 2 Within recent years, new methods in SAXS data analysis 3,4 and the introduction of automated sampling robots [5][6][7][8] have led to a rapid increase in the popularity of SAXS among structural biologists. However, while sampling robots have increased the ease of use and throughput, these robots still require 5 to 30 μl of protein volume per measurement (see Table 1).…”

Section: Introductionmentioning

confidence: 99%

LabDisk for SAXS: a centrifugal microfluidic sample preparation platform for small-angle X-ray scattering

et al. 2016

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We present a centrifugal microfluidic LabDisk for protein structure analysis via small-angle X-ray scattering (SAXS) on synchrotron beamlines. One LabDisk prepares 120 different measurement conditions, grouped into six dilution matrices. Each dilution matrix: (1) features automatic generation of 20 different measurement conditions from three input liquids and (2) requires only 2.5 μl of protein solution, which corresponds to a tenfold reduction in sample volume in comparison to the state of the art. Total hands on time for preparation of 120 different measurement conditions is less than 5 min. Read-out is performed on disk within the synchrotron beamline P12 at EMBL Hamburg (PETRA III, DESY). We demonstrate: (1) aliquoting of 40 nl aliquots for five different liquids typically used in SAXS and (2) confirm fluidic performance of aliquoting, merging, mixing and read-out from SAXS experiments (2.7-4.4% CV of protein concentration).We apply the LabDisk for SAXS for basic analysis methods, such as measurement of the radius of gyration, and advanced analysis methods, such as the ab initio calculation of 3D models. The suitability of the LabDisk for SAXS for protein structure analysis under different environmental conditions is demonstrated for glucose isomerase under varying protein and NaCl concentrations. We show that the apparent radius of gyration of the negatively charged glucose isomerase decreases with increasing protein concentration at low salt concentration. At high salt concentration the radius of gyration (R g ) does not change with protein concentrations. Such experiments can be performed by a non-expert, since the LabDisk for SAXS does not require attachment of tubings or pumps and can be filled with regular pipettes. The new platform has the potential to introduce routine high-throughput SAXS screening of protein structures with minimal input volumes to the regular operation of synchrotron beamlines.

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“…Alternatively, a custom "world-to-chip" interface consisting of a circular array of wells can programmatically infuse many reagents into a microfluidic device in a defined sequence. 12 However, the instrument must be constructed from scratch and lacks flexibility for performing other important macroscopic fluid-handling processes, like combining, purifying, and analyzing reagents.…”

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confidence: 99%

Robotic automation of droplet microfluidics

Kim

2018

Preprint

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Droplet microfluidics enables new reactions, assays, and analytic capabilities, but often requires complex workflows involving numerous steps of macro- and micro-fluidic processing. We demonstrate robotically-automated droplet microfluidics, an approach to automate workflows with commercial fluid-handling robots. These workflows can be performed without human intervention, increasing reliability and convenience.

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Automated microfluidic sample-preparation platform for high-throughput structural investigation of proteins by small-angle X-ray scattering

Cited by 33 publications

References 31 publications

In situ microfluidic dialysis for biological small-angle X-ray scattering

In situ microfluidic dialysis for biological small-angle X-ray scattering

LabDisk for SAXS: a centrifugal microfluidic sample preparation platform for small-angle X-ray scattering

Robotic automation of droplet microfluidics

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