Structure-guided disulfide engineering restricts antibody conformation to elicit TNFR agonism.

Elliott IG, Fisher H, Chan HTC, Inzhelevskaya T, Mockridge CI, Penfold CA, Duriez PJ, Orr CM, Herniman J, Müller KTJ, Essex JW, Cragg MS, Tews I, Nat Commun 16(1):3495 (2025) Europe PMC

SASDSD7 – Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S kappa chain C214S

Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S
MWexperimental 94 kDa
MWexpected 99 kDa
VPorod 124 nm3
log I(s) 1.03×102 1.03×101 1.03×100 1.03×10-1
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S small angle scattering data  s, nm-1
ln I(s)
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S Guinier plot ln 1.03×102 Rg: 3.9 nm 0 (3.9 nm)-2 s2
(sRg)2I(s)/I(0)
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S Kratky plot 1.104 0 √3 sRg
p(r)
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S pair distance distribution function Rg: 3.9 nm 0 Dmax: 13.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S kappa chain C214S Rg histogram Rg, nm
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S EOM/RANCH model
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S EOM/RANCH model
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S EOM/RANCH model
log I(s)
 s, nm-1
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S MODELLER model
log I(s)
 s, nm-1
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S PDB (PROTEIN DATA BANK) model
Synchrotron SAXS data from solutions of Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S kappa chain C214S in 50 mM HEPES, 150 mM KCl, pH 7.5 were collected on the BM29 beam line at the ESRF storage ring (Grenoble, France) using a Pilatus3 2M detector at a sample-detector distance of 2.7 m and at a wavelength of λ = 0.09918 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 45.00 μl sample at 4.5 mg/ml was injected at a 0.30 ml/min flow rate onto a Agilent Bio SEC-3, 300 Å column (300 Å pore size, 4.6 mm i.d., 300 mm length) at 20°C. 490 successive 2 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.

Full EOM/GAGOE results are made available in the full entry zip archive.

Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - heavy chain K223C/C224S
Mol. type   Protein
Olig. state   Dimer
Mon. MW   25.9 kDa
Sequence   FASTA
 
Human immunoglobulin gamma 2 (IgG2) ChiLob 7/4 F(ab')2 - kappa light chain C214S
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   23.4 kDa
Sequence   FASTA