Size-exclusion chromatography–small-angle neutron scattering system optimized for an instrument with medium neutron flux

Morishima K Inoue R, Nakagawa T, Shimizu M, Sakamoto R, Oda T, Mayumi K, Sugiyama M, Journal of Applied Crystallography 58(2) (2025) DOI

SASDWA3 – Ovalbumin, Monomer from SEC-SANS

Ovalbumin
MWI(0) 43 kDa
MWexpected 43 kDa
log I(s) 9.55×10-2 9.55×10-3 9.55×10-4 9.55×10-5
Ovalbumin small angle scattering data  s, nm-1
ln I(s)
Ovalbumin Guinier plot ln 9.55×10-2 Rg: 2.0 nm 0 (2.0 nm)-2 s2
(sRg)2I(s)/I(0)
Ovalbumin Kratky plot 1.104 0 √3 sRg
p(r)
Ovalbumin pair distance distribution function Rg: 2.1 nm 0 Dmax: 6.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Ovalbumin PDB (PROTEIN DATA BANK) model
SANS data from solutions of ovalbumin in 100 mM Tris-HCl, 100 mM NaCl in 100% v/v D₂O, pH 7.5 were collected on the SANS-U beam line at JRR-3 (Japan Atomic Energy Agency; Ibaraki, Japan) using a ORDELA, model 2660N detector at a sample-detector distance of 4 and 1.03 m at a wavelength of λ = 0.6 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 500.00 μl sample at 7.9 mg/ml was injected onto a Cytiva Superdex 200 Increase 10/300 column at 25°C. 36 successive 600 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 elution paused during SANS measurements.

Ovalbumin
Mol. type   Protein
Organism   Gallus gallus
Olig. state   Monomer
Mon. MW   42.8 kDa
 
UniProt   P01012 (2-386)
Sequence   FASTA
 
PDB ID   1OVA