TY - JOUR
T1 - Characterization of native reversible self-association of a monoclonal antibody mediated by Fab-Fab interaction
AU - Gentiluomo, Lorenzo
AU - Roessner, Dierk
AU - Streicher, Werner
AU - Mahapatra, Sujata
AU - Harris, Pernille
AU - Frieß, Wolfgang
PY - 2020
Y1 - 2020
N2 - The native reversible self-association of monoclonal antibodies has been associated with high viscosity, liquid-liquid and liquid-solid phase separation. We investigated the native reversible self-association of an IgG1, which exerts this association even at low protein concentrations, in detail to gain further understanding of this phenomenon by extensive characterization of the association as a function of multiple factors, namely pH, temperature, salt concentration and protein concentration. The nature of the self-association of the full-length IgG1 as well as the corresponding Fab and Fc fragment was studied by viz. SEC-MALS, DLS, SLS, AUC, SAXS, AF4-MALS and intrinsic fluorescence. We rationalized the self-association as a combination of hydrophobic and electrostatic interactions driven by the Fab fragments. Finally, we investigated the long-term stability of the IgG1 molecule.
AB - The native reversible self-association of monoclonal antibodies has been associated with high viscosity, liquid-liquid and liquid-solid phase separation. We investigated the native reversible self-association of an IgG1, which exerts this association even at low protein concentrations, in detail to gain further understanding of this phenomenon by extensive characterization of the association as a function of multiple factors, namely pH, temperature, salt concentration and protein concentration. The nature of the self-association of the full-length IgG1 as well as the corresponding Fab and Fc fragment was studied by viz. SEC-MALS, DLS, SLS, AUC, SAXS, AF4-MALS and intrinsic fluorescence. We rationalized the self-association as a combination of hydrophobic and electrostatic interactions driven by the Fab fragments. Finally, we investigated the long-term stability of the IgG1 molecule.
KW - Protein aggregation
KW - Protein formulation(s)
KW - Self-association
KW - Biopharmaceutical characterization
KW - Monoclonal antibody(s)
KW - Physical stability
U2 - 10.1016/j.xphs.2019.09.021
DO - 10.1016/j.xphs.2019.09.021
M3 - Journal article
C2 - 31563513
SN - 0022-3549
VL - 109
SP - 443
EP - 451
JO - Journal of Pharmaceutical Sciences
JF - Journal of Pharmaceutical Sciences
IS - 1
ER -