Identification of binding sites on human serum albumin for somapacitan - a long-acting growth hormone derivative.

Johansson E, Nielsen AD, Demuth H, Wiberg C, Schjødt C, Huang T, Chen J, Jensen S, Petersen J, Thygesen P, Biochemistry (2020) Europe PMC

SASDF32 – Ternary complex of FcRn ectodomain, FcRn binding optimised human serum albumin and the albumin-binding side chain of the human growth hormone derivative somapacitan.

FcRn binding optimised human serum albumin V418M, T420A, E505G, V547A
neonatal Fc receptor ectodomain beta-microglogulin part with C-terminal His6 tag
Albumin-binding side-chain
neonatal Fc receptor ectodomain alpha-chain
MWexperimental 110 kDa
MWexpected 111 kDa
VPorod 174 nm3
log I(s) 3.15×10-4 3.15×10-5 3.15×10-6 3.15×10-7
FcRn binding optimised human serum albumin V418M, T420A, E505G, V547A neonatal Fc receptor ectodomain beta-microglogulin part with C-terminal His6 tag Albumin-binding side-chain neonatal Fc receptor ectodomain alpha-chain small angle scattering data  s, nm-1
ln I(s)
FcRn binding optimised human serum albumin V418M, T420A, E505G, V547A neonatal Fc receptor ectodomain beta-microglogulin part with C-terminal His6 tag Albumin-binding side-chain neonatal Fc receptor ectodomain alpha-chain Guinier plot ln 3.16×10-4 Rg: 3.6 nm 0 (3.6 nm)-2 s2
(sRg)2I(s)/I(0)
FcRn binding optimised human serum albumin V418M, T420A, E505G, V547A neonatal Fc receptor ectodomain beta-microglogulin part with C-terminal His6 tag Albumin-binding side-chain neonatal Fc receptor ectodomain alpha-chain Kratky plot 1.104 0 √3 sRg
p(r)
FcRn binding optimised human serum albumin V418M, T420A, E505G, V547A neonatal Fc receptor ectodomain beta-microglogulin part with C-terminal His6 tag Albumin-binding side-chain neonatal Fc receptor ectodomain alpha-chain pair distance distribution function Rg: 3.6 nm 0 Dmax: 12.6 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of the ternary complex formed between the FcRn ectodomain, FcRn binding-optimised HSA and the albumin-binding side chain of the human growth hormone derivative somapacitan, in 100 mM MES, 100 mM NaCl, pH 6.5 were collected on the I911-4 beam line at MAX IV (Lund, Sweden) using a Pilatus 1M detector at a wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 5.00 mg/ml was measured at 25°C. One 120 second frame was collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Storage temperature = UNKNOWN. Sample detector distance = UNKNOWN

FcRn binding optimised human serum albumin V418M, T420A, E505G, V547A (HSAopt)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   66.4 kDa
 
UniProt   P02768 (25-609)
Sequence   FASTA
 
neonatal Fc receptor ectodomain beta-microglogulin part with C-terminal His6 tag (FcRn ECD beta)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   12.6 kDa
 
UniProt   P61768 (21-119)
Sequence   FASTA
 
Albumin-binding side-chain
Mol. type   Other
Olig. state   Monomer
Mon. MW   1.3 kDa
Chemical formula
 
neonatal Fc receptor ectodomain alpha-chain (FcRn ECD alpha)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Monomer
Mon. MW   30.4 kDa
 
UniProt   P55899 (24-297)
Sequence   FASTA