Characterization of native reversible self-association of a monoclonal antibody mediated by Fab-Fab interaction

Lorenzo Gentiluomo*, Dierk Roessner, Werner Streicher, Sujata Mahapatra, Pernille Harris, Wolfgang Frieß

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

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.
Original languageEnglish
JournalJournal of Pharmaceutical Sciences
Volume109
Issue number1
Pages (from-to)443-451
ISSN0022-3549
DOIs
Publication statusPublished - 2020

Keywords

  • Protein aggregation
  • Protein formulation(s)
  • Self-association
  • Biopharmaceutical characterization
  • Monoclonal antibody(s)
  • Physical stability

Cite this

Gentiluomo, Lorenzo ; Roessner, Dierk ; Streicher, Werner ; Mahapatra, Sujata ; Harris, Pernille ; Frieß, Wolfgang. / Characterization of native reversible self-association of a monoclonal antibody mediated by Fab-Fab interaction. In: Journal of Pharmaceutical Sciences. 2020 ; Vol. 109, No. 1. pp. 443-451.
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abstract = "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.",
keywords = "Protein aggregation, Protein formulation(s), Self-association, Biopharmaceutical characterization, Monoclonal antibody(s), Physical stability",
author = "Lorenzo Gentiluomo and Dierk Roessner and Werner Streicher and Sujata Mahapatra and Pernille Harris and Wolfgang Frie{\ss}",
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Characterization of native reversible self-association of a monoclonal antibody mediated by Fab-Fab interaction. / Gentiluomo, Lorenzo; Roessner, Dierk; Streicher, Werner; Mahapatra, Sujata; Harris, Pernille; Frieß, Wolfgang.

In: Journal of Pharmaceutical Sciences, Vol. 109, No. 1, 2020, p. 443-451.

Research output: Contribution to journalJournal articleResearchpeer-review

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

VL - 109

SP - 443

EP - 451

JO - Journal of Pharmaceutical Sciences

JF - Journal of Pharmaceutical Sciences

SN - 0022-3549

IS - 1

ER -