Inline small‐angle X‐ray scattering‐coupled chromatography under extreme hydrostatic pressure

Miller R, Cummings C, Huang Q, Ando N, Gillilan R, Protein Science 31(12) (2022) DOI

SASDP67 – Bovine serum albumin at neutral pH, 4 °C, and 5 MPa (1 atm) of pressure

MWexperimental 60 kDa
MWexpected 69 kDa
VPorod 99 nm3
log I(s) 3.11×100 3.11×10-1 3.11×10-2 3.11×10-3
Albumin small angle scattering data  s, nm-1
ln I(s)
Albumin Guinier plot ln 3.11×100 Rg: 2.7 nm 0 (2.7 nm)-2 s2
Albumin Kratky plot 1.104 0 3 sRg
Dmax: 8.5 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of bovine serum albumin in 25 mM HEPES, 150 mM NaCl, and 1% v/v glycerol, pH 7 were collected on the ID7A1 BioSAXS / HP-Bio Beamline beam line at the Cornell High Energy Synchrotron Source (CHESS; Ithaca, NY, USA) using a Eiger 4M detector at a sample-detector distance of 1.6 m and at a wavelength of λ = 0.089 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A sample at 20 mg/ml was injected onto a column at 4°C. 70 successive 2 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

SEC column = UNKNOWN. Sample injection volume = UNKNOWN. Flow rate = UNKNOWN

Albumin (BSA)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   69.3 kDa
UniProt   P02769 (1-607)
Sequence   FASTA