Respiratory syncytial virus (RSV)-approved monoclonal antibody Palivizumab as ligand for anti-idiotype nanobody-based synthetic cytokine receptors

Ettich J, Wittich C, Moll J, Behnke K, Floss D, Reiners J, Christmann A, Lang P, Smits S, Kolmar H, Scheller J, Journal of Biological Chemistry :105270 (2023) DOI

SASDSW6 – Palivizumab Nanobody AIP1VHH complex

Palivizumab IgG
Anti-Idiotypic-Palivizumab-Nanobody1
MWexperimental 186 kDa
MWexpected 165 kDa
log I(s) 1.70×10-1 1.70×10-2 1.70×10-3 1.70×10-4
Palivizumab IgG Anti-Idiotypic-Palivizumab-Nanobody1 small angle scattering data  s, nm-1
ln I(s)
Palivizumab IgG Anti-Idiotypic-Palivizumab-Nanobody1 Guinier plot ln 1.71×10-1 Rg: 6.0 nm 0 (6.0 nm)-2 s2
(sRg)2I(s)/I(0)
Palivizumab IgG Anti-Idiotypic-Palivizumab-Nanobody1 Kratky plot 1.104 0 √3 sRg
p(r)
Palivizumab IgG Anti-Idiotypic-Palivizumab-Nanobody1 pair distance distribution function Rg: 6.2 nm 0 Dmax: 20.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Palivizumab IgG Anti-Idiotypic-Palivizumab-Nanobody1 CORAL model
Synchrotron SAXS data from solutions of Palivizumab Nanobody AIP1VHH complex in PBS (137 mM NaCl, 2.7 mM KCl, 12 mM HPO4 2−/H2PO4 −), pH 7.4 were collected on the EMBL P12 beam line at PETRA III (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 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 100.00 μl sample at 5.5 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 10°C. 3000 successive 0.995 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.

Palivizumab IgG (Palivizumab)
Mol. type   Protein
Organism   Commercial (Synagis)
Olig. state   Monomer
Mon. MW   144.9 kDa
Sequence   FASTA
 
Anti-Idiotypic-Palivizumab-Nanobody1 (AIP1VHH)
Mol. type   Protein
Organism   Lama glama
Olig. state   Monomer
Mon. MW   20.2 kDa
Sequence   FASTA