Insulin Binding to the Analytical Antibody Sandwich Pair OXI ‐005 and HUI ‐018 – Epitope Mapping and Binding Properties

Johansson E, Wu X, Yu B, Yang Z, Cao Z, Wiberg C, Jeppesen C, Poulsen F, Protein Science (2020) DOI

SASDJ24 – Immunoglobulin HUI-018 Fab and OXI-005 Fab in a ternary complex with porcine insulin

HUI-018 Fab
Insulin (Insulin B chain and Insulin A chain)
OXI-005 Fab
MWexperimental 106 kDa
MWexpected 100 kDa
VPorod 138 nm3
log I(s) 1.50×101 1.50×100 1.50×10-1 1.50×10-2
HUI-018 Fab Insulin (Insulin B chain and Insulin A chain) OXI-005 Fab small angle scattering data  s, nm-1
ln I(s)
HUI-018 Fab Insulin (Insulin B chain and Insulin A chain) OXI-005 Fab Guinier plot ln 1.51×101 Rg: 4.5 nm 0 (4.5 nm)-2 s2
(sRg)2I(s)/I(0)
HUI-018 Fab Insulin (Insulin B chain and Insulin A chain) OXI-005 Fab Kratky plot 1.104 0 3 sRg
p(r)
HUI-018 Fab Insulin (Insulin B chain and Insulin A chain) OXI-005 Fab pair distance distribution function Rg: 4.6 nm 0 Dmax: 14.2 nm

Data validation


Fits and models


log I(s)
 s, nm-1
HUI-018 Fab Insulin (Insulin B chain and Insulin A chain) OXI-005 Fab OTHER model

SAXS data from solutions of HUI-018 Fab and OXI-005 Fab in a ternary complex with porcine insulin in 20 mM HEPES, 140 mM NaCl, pH 7.5 were collected using a Rigaku BioSAXS-2000 equipped with a Pilatus 100K detector (Novo Nordisk A/S, Maaloev, Denmark) at a wavelength of λ = 0.15418 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 2.00 mg/ml was measured at 20°C. Six 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.

Sample detector distance = UNKNOWN

HUI-018 Fab (HUI-018Fab)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   47.5 kDa
Sequence   FASTA
 
Insulin (Insulin B chain and Insulin A chain) (INS)
Mol. type   Protein
Organism   Sus scrofa
Olig. state   Monomer
Mon. MW   5.8 kDa
 
UniProt   P01315 (25-108)
Sequence   FASTA
 
OXI-005 Fab (OXI-005Fab)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   47.2 kDa
Sequence   FASTA