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HP-BioSANS (LIPSS)

LIPSS Schematics

High-Pressure BioSANS

LIQUID INSERTION PRESSURE SYSTEM FOR SANS (LIPSS)

LIPSS enables in situ SANS measurements under broad ranges of pressure and temperature, so that sample environment effects can be probed from minutes to hours1. No radiation damage and an extended q-range make HP-SANS complementary to HP-SAXS measurements, providing information on a broad range of molecular sizes, shape, aggregation, folding/unfolding, etc. The non destructive nature of neutrons allows for hysteresis effects to be probed, while contrast variation SANS can be used to highlight contributions to scattering from specific components of complex macromolecular assemblies such as viruses2 or others.

LIPSS is a high-pressure (HP) system (see specifications below): the sample cell is made of a high-nickel-content austenitic steel and is capable of withstanding a maximum pressure up to 350 MPa (3.5 kbar). A Peltier system controls the temperature between -20°C and +65°C: under pressure, subzero temperatures can be accessed in the absence of ice. The sample is loaded through an injection system that minimizes air bubbles and fills the cell completely. A separator isolates the sample from the pressurizing medium, as shown schematically in the image above1. LIPSS was designed for low viscosity solutions and its performance depends on the specific environment required. If you are interested in using LIPSS it is strongly recommended that you discuss measurement needs in advance with Susana Teixeira [scm5(at)nist.gov] .

Past and potential applications of HP-SANS and the LIPSS sample environment include studies on:

  • cold denaturation of proteins3
  • subzero temperature effects on storage of monoclonal antibodies4
  • high-pressure low-temperature processing of food proteins5
  • high-pressure response of amyloid folds6
  • self-assembled lipid nanoparticles7
  • viral inactivation8,9
  • pressure-assisted enzymatic digestion of antibodies10
  • protein aggregation under pressure11,12
  • lipid phase transitions13
  • surfactant self-assembly at high pressure14
  • polymer blend nucleation and interactions15,16
  • engineering baroplastic behavior of block copolymers17

REFERENCES

  1. S. Teixeira et al., High Pressure Cell for Bio-SANS Studies Under Sub-zero Temperatures or Heat Denaturing Conditions’. J. Neutron Res 20,13 – 23 (2018).
  2. L. He et al., Conformational changes in Sindbis virus induced by decreased pH are revealed by small-angle neutron scattering. J Virol 86, 1982-1987 (2012)
  3. C. Dias et al., The hydrophobic effect and its role in cold denaturation, Cryobiology 60 ,91–99 (2010).
  4. K. Lazar et al., Cold denaturation of monoclonal antibodies. mAbs 2, 42 - 52 (2010).
  5. S.C.M Teixeira. High-pressure small-angle neutron scattering for food studies, Curr. Op. Colloid & Interface Science 42, 99-109 (2019).
  6. J. Torrent et al., High pressure response of amyloid folds. Viruses 11(3): 202 (2019).
  7. C. Kulkarni et al., Effects of High Pressure on Internally Self-Assembled Lipid Nanoparticles: A Synchrotron Small-Angle X-ray Scattering (SAXS) Study. Langmuir 32, 45, 11907–11917 (2016).
  8. Oliveira et al., Low Temperature and Pressure Stability of Picornaviruses: Implications for Virus Uncoating. Biophysical J. 76, 3, 1270-1279 (1999).

  9. D. Kingsley. High Pressure Processing and its Application to the Challenge of Virus-Contaminated Foods. Food Environ Virol. 5(1): 1–12 (2013).

  10. Fang et al. (2016) Pressurized online pepsin digestion of mAb IgG2 for Hydrogen Deuterium Exchange Mass Spectrometry. https://www.waters.com/webassets/cms/library/docs/720005693en.pdf. Accessed April 2020.

  11. Jackson & McGillivray. Protein aggregate structure under high pressure. Chem. Commun. 47, 487–489 (2011).

  12. Seefeldt et al., High-pressure studies of aggregation of recombinant human interleukin-1 receptor antagonist: Thermodynamics, kinetics, and application to accelerated formulation studies. Protein Science 14(9): 2258–2266 (2005).

  13. Hammouda and Clover. SANS from P85/Water-d under Pressure. Langmuir 26(9), 6625–6629 (2010).

  14. Leseman et al., Self-Assembly at High Pressures: SANS Study of the Effect of Pressure on Microstructure of C8E5 Micelles in Water. Ind. Eng. Chem. Res. 42, 6425-6430 (2003).

  15. Patel et al., Observing Nucleation Close to the Binodal by Perturbing Metastable Polymer Blends. Macromol. 40(5), 1675 (2007).

  16. Ruegg et al., Effect of Pressure on a Multicomponent A/B/A-C Polymer Blend with Attractive and Repulsive Interactions. Macromol. 40(2), 355 (2007).

  17. Ruzette et al., Pressure effects on the phase behavior of styrene/n-alkyl methacrylate block copolymers. Macromol. 36(9), 3351 (2003).

USEFUL LINKS 

Specifications/Capabilities

  • Pressure range (MPa):                           0.1 - 350  (ambient to 3.5 kbar)
  • Temperature range (°C):                       -20 to +65 (higher temperatures possible§)
  • Sample volume (mL):                               2 to 5
  • Typical sample concentration:             5 mg/mL or higher (depends on contrast§)
  • Can I recover my sample?                   Yes, typical recovery of ~2/3 of the volume
  • Sample thickness (mm):                 1 to 5 (neutron path length across the sample solution)
  • Q range (Å-1):                                    0.003 to 0.3 (SANS); 0.0001 to 0.003 (USANS BT5 instrument)
  • Type of samples:                                   Liquid (solutions or stable suspensions)
  • Real space distances:                      ~30 Å up to ~6 µm (depends on resolution and contrast§)
  • Training required?                            Yes, please contact S. Teixeira  or J. Leão to arrange for training in advance.

Compatible with NCNR instruments: NGB30SANS, USANS (BT5), NG7SANS, 10m SANS.

§ Please contact Susana Teixeira or your local contact if you have questions regarding feasibility.

Scientific Opportunities/Applications

Created July 6, 2020, Updated July 7, 2020