Size-exclusion chromatography small-angle X-ray scattering of water soluble proteins on a laboratory instrument.

Bucciarelli S, Midtgaard SR, Nors Pedersen M, Skou S, Arleth L, Vestergaard B, J Appl Crystallogr 51(Pt 6):1623-1632 (2018) Europe PMC

SASDE95 – Bovine serum albumin monomer from in-house SEC-SAXS

Bovine serum albumin
MWexperimental 61 kDa
MWexpected 66 kDa
VPorod 74 nm3
log I(s) 6.90×10-2 6.90×10-3 6.90×10-4 6.90×10-5
Bovine serum albumin small angle scattering data  s, nm-1
ln I(s)
Bovine serum albumin Guinier plot ln 6.90×10-2 Rg: 2.9 nm 0 (2.9 nm)-2 s2
(sRg)2I(s)/I(0)
Bovine serum albumin Kratky plot 1.104 0 3 sRg
p(r)
Bovine serum albumin pair distance distribution function Rg: 2.8 nm 0 Dmax: 8.6 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Bovine serum albumin PDB (PROTEIN DATA BANK) model

log I(s)
 s, nm-1
Bovine serum albumin DAMFILT model

SAXS data from solutions of Bovine serum albumin monomer from in-house SEC-SAXS in PBS, pH 7.4 were collected on a Xenocs BioXolver L instrument at the Copenhagen University, Department of Drug Design and Pharmacology (Copenhagen, Denmark) using a 20Hz Pilatus 300K detector at a sample-detector distance of 0.7 m and at a wavelength of λ = 0.134 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle).

The BSA sample (500 microlitres at 1.0 mg/ml) was injected onto a GE Healthcare S200 Increase 10/300 (24 ml) column at a flow rate of 0.1 ml/min at 23°C. A total of 100 x 30 second SAXS data frames were recorded throughout the BSA elution. After background solvent corrections, SEC-SAXS data frames 60-69, corresponding to monomeric BSA, were scaled, averaged and binned logarithmically to produce the data displayed in this entry. Additional files, including plots of the selected frames and of I(0), HPLC and exposure cell UV traces and Rg vs. time (determined by AUTORG) are included in the additional files of the full entry zip archive.

Bovine serum albumin (BSA)
Mol. type   Protein
Organism   Bos taurus
Olig. state   Monomer
Mon. MW   66.4 kDa
 
UniProt   P02769 (25-607)
Sequence   FASTA
 
PDB ID   4F5S