Behavior of Monoclonal Antibodies: Relation Between the Second Virial Coefficient (B 2) at Low Concentrations and Aggregation Propensity and Viscosity at High Concentrations

Saito, Shuntaro; Hasegawa, Jun; Kobayashi, Naoki; Kishi, Naoyuki; Uchiyama, Susumu; Fukui, Kiichi

doi:10.1007/s11095-011-0563-x

Cited by 133 publications

(140 citation statements)

References 42 publications

(60 reference statements)

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“…More importantly, we have provided examples where aggregation does not correlate with conformational stability and native-state protein-protein interactions. As such, there will be limited utility of using k D values as a predictor for aggregate growth rates in the absence of strong electrostatic interactions [7,78].…”

Section: Discussionmentioning

confidence: 99%

“…A decreasing k D value is often correlated with increased aggregation propensity when reducing electrostatic repulsion by increasing ionic strength or using a salt or buffer ion that neutralizes protein charge [35,46,57,74,78,82,19,46,63,77,81]. Consistent with these studies, for the wild-type, SB and DSV mutants, the relative rates of aggregate growth (as reflected by the corresponding T SLS ⁄ values) and protein-protein interactions (as reflected by the k D values) decrease non-specifically upon addition of NaCl due to screening of electrostatic interactions.…”

Section: Correlations Between K D and Aggregation Propensitymentioning

confidence: 99%

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The effect of charge mutations on the stability and aggregation of a human single chain Fv fragment

Austerberry

Dajani

Panova

et al. 2017

European Journal of Pharmaceutics and Biopharmaceutics

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a b s t r a c tThe aggregation propensities for a series of single-chain variable fragment (scFv) mutant proteins containing supercharged sequences, salt bridges and lysine/arginine-enriched motifs were characterised as a function of pH and ionic strength to isolate the electrostatic contributions. Recent improvements in aggregation predictors rely on using knowledge of native-state protein-protein interactions. Consistent with previous findings, electrostatic contributions to native protein-protein interactions correlate with aggregate growth pathway and rates. However, strong reversible self-association observed for selected mutants under native conditions did not correlate with aggregate growth, indicating 'sticky' surfaces that are exposed in the native monomeric state are inaccessible when aggregates grow. We find that even though similar native-state protein-protein interactions occur for the arginine and lysine-enriched mutants, aggregation propensity is increased for the former and decreased for the latter, providing evidence that lysine suppresses interactions between partially folded states under these conditions. The supercharged mutants follow the behaviour observed for basic proteins under acidic conditions; where excess net charge decreases conformational stability and increases nucleation rates, but conversely reduces aggregate growth rates due to increased intermolecular electrostatic repulsion. The results highlight the limitations of using conformational stability and native-state protein-protein interactions as predictors for aggregation propensity and provide guidance on how to engineer stabilizing charged mutations.

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Section: Discussionmentioning

confidence: 99%

Section: Correlations Between K D and Aggregation Propensitymentioning

confidence: 99%

The effect of charge mutations on the stability and aggregation of a human single chain Fv fragment

Austerberry

Dajani

Panova

et al. 2017

European Journal of Pharmaceutics and Biopharmaceutics

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“…Saito et al 38 suggested the following general guideline: Antibodies with B 22 greater than 2£10 ¡5 mL mol per gm 2 have solution viscosities lower than 20 cP at 150 mg per mL, provided B 22 and viscosity measurements are performed in the same formulation. 38 In a separate study, Connolly et al 39 40 In the data from Binabaji et al, mAbs that met the B 22 criterion of Saito et al had low solution viscosities at 150 mg per mL. However, Binabaji et al also reported that B 22 is insufficient to predict solution viscosities when mAb concentrations rise above 200 mg per mL.…”

Section: Theoretical Foundations and Experimentally Measurable Quantimentioning

confidence: 99%

Molecular basis of high viscosity in concentrated antibody solutions: Strategies for high concentration drug product development

Tomar

Kumar

Singh

et al. 2016

mAbs

153

145

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Effective translation of breakthrough discoveries into innovative products in the clinic requires proactive mitigation or elimination of several drug development challenges. These challenges can vary depending upon the type of drug molecule. In the case of therapeutic antibody candidates, a commonly encountered challenge is high viscosity of the concentrated antibody solutions. Concentration-dependent viscosity behaviors of mAbs and other biologic entities may depend on pairwise and higher-order intermolecular interactions, non-native aggregation, and concentration-dependent fluctuations of various antibody regions. This article reviews our current understanding of molecular origins of viscosity behaviors of antibody solutions. We discuss general strategies and guidelines to select low viscosity candidates or optimize lead candidates for lower viscosity at early drug discovery stages. Moreover, strategies for formulation optimization and excipient design are also presented for candidates already in advanced product development stages. Potential future directions for research in this field are also explored.

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“…Herein, B 2 was determined from three concentrations less than 10 mg/mL in solutions of varying pH for three different MAbs. All MAbs, MAb-A, MAb-B, and MAb-C, are humanized IgG 1 which have the same constant region and different variable regions (Saito et al 2012). B 2 determined by AUC-SE at different pH values is shown in Fig.…”

Section: Predictions Of Aggregation Propensity and Viscosity At High mentioning

confidence: 99%

Biopharmaceutical Evaluation of Intermolecular Interactions by AUC-SE

Saito

Uchiyama

2016

Analytical Ultracentrifugation

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Analytical ultracentrifugation sedimentation equilibrium (AUC-SE) is a useful technique to investigate the weak reversible intermolecular interactions among proteins in solution. It provides biophysical information such as the average apparent molecular weight, stoichiometry, and association constant of associating proteins. Several studies of intermolecular interaction in biopharmaceuticals by AUC-SE are introduced in this chapter. AUC-SE also provides the second virial coefficient (B 2 ), which represents the type, i.e., repulsive and attractive, and a magnitude of intermolecular interactions. The B 2 values obtained from the protein solution at low concentrations showed good correlation with aggregation and viscosity of MAb at high concentrations, indicating that B 2 can be an effective indicator of aggregation propensity and viscosity. These findings suggest that AUC-SE provides clues to understand the self-association in biopharmaceuticals and to establish effective manufacturing process, formulation, and administration of biopharmaceuticals.

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Behavior of Monoclonal Antibodies: Relation Between the Second Virial Coefficient (B 2) at Low Concentrations and Aggregation Propensity and Viscosity at High Concentrations

Abstract: B (2) can be used, within certain limitations, as an effective indicator of viscosity and aggregation propensity of highly concentrated MAb solutions.

Cited by 133 publications

References 42 publications

The effect of charge mutations on the stability and aggregation of a human single chain Fv fragment

The effect of charge mutations on the stability and aggregation of a human single chain Fv fragment

Molecular basis of high viscosity in concentrated antibody solutions: Strategies for high concentration drug product development

Biopharmaceutical Evaluation of Intermolecular Interactions by AUC-SE

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