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Title: Analysis of MonoPEGylated human galectin-2 by small-angle x-ray and neutron scattering: concentration dependence of PEG conformation in the conjugate.
Authors: He, LZ
Wang, H
Garamus, VM
Hanley, T
Lensch, M
Gabius, HJ
Fee, CJ
Middelberg, A
Keywords: Small Angle Scattering
Polyethylene Glycols
Abundance
Proteins
Molecules
Agglomeration
Issue Date: 1-Dec-2010
Publisher: American Chemical Society
Citation: He, L. Z., Wang, H., Garamus, V. M., Hanley, T., Lensch, M., Gabius, H. J., et al. (2010). Analysis of MonoPEGylated human galectin-2 by small-angle x-ray and neutron scattering: concentration dependence of PEG conformation in the conjugate. Biomacromolecules, 11(12), 3504-3510.
Abstract: Protein conjugation with polyethylene glycol (PEG) is a valuable means for improving stability, solubility, and bioavailability of pharmaceutical proteins. Using human galectin-2 (hGal-2) and 5 kDa PEG as a model system we first produced a PEG-hGal-2 conjugate exclusively at the Cys75 residue, resulting in two monosubstituted subunits per hGal-2 homodimer. Small angle X-ray and neutron scattering (SAXS and SANS) were combined to provide complementary structural information about the PEG-hGal-2 conjugate, wherein signal generation in SAXS depends mainly on the protein while SANS data presents signals from both the protein and PEG moieties. SAXS data gave a constant radius of gyration (Rg = 21.5 Å) for the conjugate at different concentrations and provided no evidence for an alteration of homodimeric structure or hGal-2 ellipsoidal shape upon PEGylation. In contrast, SANS data revealed a concentration dependence of Rg for the conjugate, with the value decreasing from 31.5 Å at 2 mg/mL to 26 Å at 14 mg/mL (based on hGal-2 concentration). Scattering data have been successfully described by the model of the ellipsoidal homogeneous core (hGal-2) attached with polymer chains (PEG) at the surface. Evidently, the PEG conformation of the conjugate strongly depends on conjugate concentration and PEG’s radius of gyration decreases from 24.5 to 15 Å. An excluded volume effect, arising from steric clashes between PEG molecules at high concentration, was quantified by estimating the second virial coefficient, A2, of PEGylated hGal-2 from the SANS data. A positive value of A2 (6.0 ± 0.4 × 10−4 cm3 mol g−2) indicates repulsive interactions between molecules, which are expected to protect the PEGylated protein against aggregation. © 2010, American Chemical Society
URI: http://dx.doi.org/10.1021/bm100999a
http://apo.ansto.gov.au/dspace/handle/10238/2932
ISSN: 1525-7797
Appears in Collections:Journal Articles

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