Improving Viscosity and Stability of a Highly Concentrated Monoclonal Antibody Solution with Concentrated Proline

Abstract

Proline demonstrated greater efficacy for improving mAb viscosity and stability in contrast to glycine and trehalose due to its amphipathic structure and partial charge on the pyrrolidine side chain. These properties likely allow proline to screen the attractive electrostatic and hydrophobic interactions that promote self-association and high viscosities. Binary proline-histidine formulations also demonstrated greater viscosity reduction effects than histidine alone at the same total co-solute concentration, while maintaining a lower total solution osmolarity.

Characterization of Excipient Effects on Reversible Self-Association, Backbone Flexibility, and Solution Properties of an IgG1 Monoclonal Antibody at High Concentrations: Part 1

“…76 Also, Arg has a relatively larger surface area than Lys, which contributes more steric repulsion, leading to fewer surface-exposed attraction sites on mAb-J and thus might reduce the chance for protein interactions. 69 Other studies also indicate Arg can increase protein solubility, increase second osmotic virial coefficient (B 22 , with increasing positive values indicating repulsive effects), and suppress formation of protein dimers, which are consistent with our results (Figs. 4 and 5).…”

“…68 Also, NaCl at low concentrations does not have an ability to modify hydrophobic interactions. 69,70 In summary, there are more possible noncovalent interactions between the mAb-J and charged amino acids than between mAb-J and NaCl, and thus, charged amino acids more effectively disrupted RSA at high protein concentrations.…”

Physicochemical Stability of Monoclonal Antibodies: A Review

“…156 Proline is a cyclic amino acid that is also claimed to enhance mAb solubilization by binding to aromatic residues and exposed hydrophobic regions and thus decreasing protein-protein interactions and solution viscosity, reducing aggregation at pH near their pI. 157,158 Glycine is used as a bulking agent in freezedried formulations. 148 Like mannitol, even if used for mechanical properties of the lyophilized formulations, it may also possess stabilization properties toward protein aggregation if remaining in amorphous state, being then more protective.…”

Molecular computations of preferential interactions of proline, arginine.HCl, and NaCl with IgG1 antibodies and their impact on aggregation and viscosity

“…The preferential interaction coefficient of antibodyexcipient mixtures can be measured experimentally via vapor pressure osmometry, 15 and is also accessible via simulation. 11 Numerous experimental [16][17][18][19] and simulation 6,7,[20][21][22][23] studies have been performed on the effects of amino acid and ionic excipients on proteins. However, many of the studies that have been performed focus on how excipients interact with only one or two proteins.…”

“…3,[24][25][26] This study aims to examine the connection between antibody surface properties and excipient interactions. While trends in the impact of certain ionic and amino acid excipients on mAbs have been studied experimentally, 16,17,18,19 and trends in the impact of antibody sequences and structures have been studied, 3,25,27 the molecular mechanisms behind the impact of excipients on mAb behavior are not well understood. Here, we use simulations and experiments to investigate the mechanisms by which three common amino acid and ionic excipients (proline, Arg.HCl, and NaCl) affect the aggregation and viscosity of three antibodies studied previously.…”