Small-angle neutron scattering characterization of monoclonal antibody conformations and interactions at high concentrations.
Small-angle neutron scattering (SANS) is used to probe the solution structure of two protein therapeutics (monoclonal antibodies 1 and 2 (MAb1 and MAb2)) and their protein-protein interaction (PPI) at high concentrations. These MAbs differ by small sequence alterations in the complementarity-determining region but show very large differences in solution viscosity. The analyses of SANS patterns as a function of different solution conditions suggest that the average intramolecular structure of both MAbs in solution is not significantly altered over the studied protein concentrations and experimental conditions. Even though a strong repulsive interaction is expected for both MAbs due to their net charges and low solvent ionic strength, analysis of the SANS data shows that the effective PPI for MAb1 is dominated by a very strong attraction at small volume fraction that becomes negligible at large concentrations. The MAb1 PPI cannot be modeled simply by a spherically symmetric central forces model. It is proposed that an anisotropic attraction strongly affects the local interprotein structure and leads to an anomalously large viscosity of concentrated MAb1 solutions. Conversely, MAb2 displays a repulsive interaction potential throughout the concentration series probed and a comparatively small solution viscosity.
Department of Chemical and Biomolecular Engineering, Center for Neutron Science, University of Delaware, Newark, USA.
Eric J
EJ
Yearley
Isidro E
IE
Zarraga
Steven J
SJ
Shire
Thomas M
TM
Scherer
Yatin
Y
Gokarn
Norman J
NJ
Wagner
Yun
Y
Liu
D000818:Animals; D016880:Anisotropy; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry* / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D033363:Neutron Diffraction; D009994:Osmolar Concentration; D011485:Protein Binding; D011487:Protein Conformation; D053838:Scattering, Small Angle; D012996:Solutions / Q000737:chemistry; D014783:Viscosity
D061067:Antibodies, Monoclonal, Humanized; D012996:Solutions