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J Pharm Sci. 2019 Jun 10. pii: S0022-3549(19)30368-5. doi: 10.1016/j.xphs.2019.06.001. [Epub ahead of print]

Characterization of Excipient Effects on Reversible Self-association, Backbone Flexibility, and Solution Properties of an IgG1 Monoclonal Antibody at High Concentrations: Part 2.

Author information

1
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, USA, 66047; Vaccine Analytics and Formulation Center, University of Kansas, Lawrence, Kansas, USA, 66047.
2
Department of Formulation Sciences, MedImmune LLC, Gaithersburg, Maryland, USA 20878.
3
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, USA, 66047.
4
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, USA, 66047; Vaccine Analytics and Formulation Center, University of Kansas, Lawrence, Kansas, USA, 66047. Electronic address: volkin@ku.edu.
5
Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, Kansas, USA, 66047; Department of Chemistry and R.N. Adams Institute of Bioanalytical Chemistry, University of Kansas, Lawrence, Kansas, USA, 66045. Electronic address: dweis@ku.edu.

Abstract

In this work, we continue to examine excipient effects on the reversible self-association (RSA) of two different IgG1 monoclonal antibodies (mAb-J and mAb-C). We characterize the RSA behavior of mAb-C which, similar to mAb-J (see companion paper), undergoes concentration-dependent RSA, but by a different molecular mechanism. Five additives that affect protein hydrophobic interactions to varying extents including a chaotropic salt (guanidine hydrochloride, GdnHCl), a hydrophobic salt (trimethylphenylammonium iodide, TMPAI), an aromatic amino acid derivative (tryptophan amide hydrochloride, TrpNH2HCl), a kosmotropic salt (sodium sulfate, Na2SO4), and a less polar solvent (ethanol) were evaluated to determine their effects on the solution properties, molecular properties, and RSA of mAb-C at various protein concentrations. Four of the five additives examined demonstrated favorable effects on the pharmaceutical properties of high concentration mAb-C solutions (i.e., lower viscosity and weakened protein-protein interactions, PPIs) with a ranking order of GdnHCl > TMPAI > TrpNH2HCl > ethanol as measured by various biophysical techniques. Conversely, addition of Na2SO4 resulted in less desirable solution properties and enhanced PPIs. The effect of these five additives on mAb-C backbone dynamics were evaluated by HX-MS (at high vs. low protein concentrations) to better understand their effects on the molecular sites of RSA in mAb-C.

KEYWORDS:

Monoclonal antibody; colloidal stability; excipient effects; hydrogen exchange-mass spectrometry; local dynamics; phase separation; protein-protein interactions; reversible self-association

PMID:
31195015
DOI:
10.1016/j.xphs.2019.06.001

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