Contrasting the Influence of Cationic Amino Acids on the Viscosity and Stability of a Highly Concentrated Monoclonal Antibody

Dear, Barton J.; Hung, Jessica; Truskett, Thomas M.; Johnston, Keith P.

doi:10.1007/s11095-016-2055-5

Cited by 54 publications

(79 citation statements)

References 54 publications

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“…where c and d are adjustable parameters. This functional form is suggested by the experimental observations [34, 50], indicating a strong increase of solution viscosity for mAb concentrations above 90 mg/mL. Note that f ( n ) depends solely on the number of molecules involved, n , and not on their spatial distribution in the cluster.…”

Section: Solution Viscosity Depends On the Antibody Cluster–size Dmentioning

confidence: 55%

Controlling the viscosities of antibody solutions through control of their binding sites

Kastelic

Dill

Kalyuzhnyi

et al. 2018

Journal of Molecular Liquids

110

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For biotechnological drugs, it is desirable to formulate antibody solutions with low viscosities. We go beyond previous colloid theories in treating protein–protein self–association of molecules that are antibody–shaped and flexible and have spatially specific binding sites. We consider interactions either through fragment antigen (Fab–Fab) or fragment crystalizable (Fab–Fc) binding. Wertheim's theory is adapted to compute the cluster–size distributions, viscosities, second virial coefficients, and Huggins coefficients, as functions of antibody concentration. We find that the aggregation properties of concentrated solutions can be anticipated from simpler–to–measure dilute solutions. A principal finding is that aggregation is controllable, in principle, through modifying the antibody itself, and not just the solution it is dissolved in. In particular: (i) monospecific antibodies having two identical Fab arms can form linear chains with intermediate viscosities. (ii) Bispecific antibodies having different Fab arms can, in some cases, only dimerize, having low viscosities. (iii) Arm–to–Fc binding allows for three binding partners, leading to networks and high viscosities.

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Section: Solution Viscosity Depends On the Antibody Cluster–size Dmentioning

confidence: 55%

Controlling the viscosities of antibody solutions through control of their binding sites

Kastelic

Dill

Kalyuzhnyi

et al. 2018

Journal of Molecular Liquids

110

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“…Nevertheless, depending on temperature, the type of salt (Hofmeister series), salt concentration, and protonation state of the protein (net positive charge at pH < pI; net neutral charge at pHp I; net negative charge at pH > pI), complex scenarios of solubility reduction and solubility enhancement ("salting in") have been reported. [89][90][91][92][93][94] The addition of (chaotropic) salts 86,95,96 or charged excipients, for example, amino acids with a hydrophobic moiety such as histidine and arginine 96,97 are reported to lower the tendency to form associates, which in turn also lowers the viscosity of the solution. Similarly, reversible association that leads to liquidliquid phase separation was recently reported to follow similar trends 92,[98][99][100][101] ; this is seen especially at low temperatures, for example, during intermediate storage during DSP.…”

Section: Purification/chromatographic Steps: Impact Of Salt and Ionicmentioning

confidence: 99%

Stress Factors in mAb Drug Substance Production Processes: Critical Assessment of Impact on Product Quality and Control Strategy

Das

Narhi

Sreedhara³

et al. 2020

Journal of Pharmaceutical Sciences

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The success of biotherapeutic development heavily relies on establishing robust production platforms. During the manufacturing process, the protein is exposed to multiple stress conditions that can result in physical and chemical modifications. The modified proteins may raise safety and quality concerns depending on the nature of the modification. Therefore, the protein modifications potentially resulting from various process steps need to be characterized and controlled. This commentary brings together expertise and knowledge from biopharmaceutical scientists and discusses the various manufacturing process steps that could adversely impact the quality of drug substance (DS). The major process steps discussed here are commonly used in mAb production using mammalian cells. These include production cell culture, harvest, antibody capture by protein A, virus inactivation, polishing by ion-exchange chromatography, virus filtration, ultrafiltration-diafiltration, compounding followed by release testing, transportation and storage of final DS. Several of these process steps are relevant to protein DS production in general. The authors attempt to critically assess the level of risk in each of the DS processing steps, discuss strategies to control or mitigate protein modification in these steps, and recommend mitigation approaches including guidance on development studies that mimic the stress induced by the unit operations.

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“…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.…”

Section: Introductionmentioning

confidence: 99%

“…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.…”

Section: Introductionmentioning

confidence: 99%

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

Cloutier

Sudrik

Mody

et al. 2020

mAbs

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Preferential interactions of excipients with the antibody surface govern their effect on the stability of antibodies in solution. We probed the preferential interactions of proline, arginine.HCl (Arg.HCl), and NaCl with three therapeutically relevant IgG1 antibodies via experiment and simulation. With simulations, we examined how excipients interacted with different types of surface patches in the variable region (Fv). For example, proline interacted most strongly with aromatic surfaces, Arg.HCl was included near negative residues, and NaCl was excluded from negative residues and certain hydrophobic regions. The differences in interaction of different excipients with the same surface patch on an antibody may be responsible for variations in the antibody's aggregation, viscosity, and self-association behaviors in each excipient. Proline reduced self-association for all three antibodies and reduced aggregation for the antibody with an association-limited aggregation mechanism. The effects of Arg.HCl and NaCl on aggregation and viscosity were highly dependent on the surface charge distribution and the extent of exclusion from highly hydrophobic patches. At pH 5.5, both tended to increase the aggregation of an antibody with a strongly positive charge on the Fv, while only NaCl reduced the aggregation of the antibody with a large negative charge patch on the Fv. Arg.HCl reduced the viscosities of antibodies with either a hydrophobicity-driven mechanism or a charge-driven mechanism. Analysis of this data presents a framework for understanding how amino acid and ionic excipients interact with different protein surfaces, and how these interactions translate to the observed stability behavior.

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Contrasting the Influence of Cationic Amino Acids on the Viscosity and Stability of a Highly Concentrated Monoclonal Antibody

Cited by 54 publications

References 54 publications

Controlling the viscosities of antibody solutions through control of their binding sites

Controlling the viscosities of antibody solutions through control of their binding sites

Stress Factors in mAb Drug Substance Production Processes: Critical Assessment of Impact on Product Quality and Control Strategy

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

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