The quaternary structure of insulin glargine and glulisine under formulation conditions.

Nagel N, Graewert MA, Gao M, Heyse W, Jeffries CM, Svergun D, Berchtold H, Biophys Chem 253:106226 (2019) Europe PMC

SASDFA4 – Insulin glargine (Toujeo®), oligomeric composition

Insulin glargine (Toujeo®)
MWI(0) 35 kDa
MWexpected 36 kDa
log I(s) 6.58×102 6.58×101 6.58×100 6.58×10-1
Insulin glargine (Toujeo®) small angle scattering data  s, nm-1
ln I(s)
Insulin glargine (Toujeo®) Guinier plot ln 6.58×102 Rg: 1.8 nm 0 (1.8 nm)-2 s2
(sRg)2I(s)/I(0)
Insulin glargine (Toujeo®) Kratky plot 1.104 0 3 sRg
p(r)
Insulin glargine (Toujeo®) pair distance distribution function Rg: 1.9 nm 0 Dmax: 6.2 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Insulin glargine (Toujeo®) PDB (PROTEIN DATA BANK) model
Insulin glargine (Toujeo®) PDB (PROTEIN DATA BANK) model

log I(s)
 s, nm-1

log I(s)
 s, nm-1

Synchrotron SAXS data from solutions of Insulin glargine (Toujeo®), oligomeric composition in Toujeo Fromulation (190 ug Zinc chloride, 2.7 mg m-Cresol, 20 mg glycerol 85%), pH 4 were collected on the EMBL P12 beam line at the PETRA III storage ring (Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 1.241 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 10.91 mg/ml was measured at 20°C. 20 successive 0.045 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.

Here, Toujeo measured at 10.91 mg/ml is displayed (formulation concentration). Measurements from 2 dilutions have been deposited in addition. Mixture analysis of the scattering profiles was performed with the program Oligomer. Data can be fit with a stable equilibrium of ~ 60 % hexamers (based on 1ev3.pdb) and ~ 40 % dimers. For higher concentration a strong influence of interparticle interference is observed, so that data at low s (>0.05 nm-1) is omitted for the fit. The presence of 3% dodecamers (based on 3W80.pdb) slightly improves the fit.

Insulin glargine (Toujeo®)
Mol. type   Protein
Olig. state   Hexamer
Mon. MW   6.1 kDa
Sequence   FASTA
 
PDB ID   1EV3
 
PDB ID   1EV3