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