Relevant shaking stress conditions for antibody preformulation development

Eppler, Annette; Weigandt, Markus; Hanefeld, Andrea; Bunjes, Heike

doi:10.1016/j.ejpb.2009.11.005

Cited by 47 publications

(37 citation statements)

References 23 publications

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“…Semiautomated visual inspection has been used to roughly detect differences in protein particles generated by several stress types in terms of number and size, thereby complementing light obscuration and turbidity results 33. Furthermore, additional phenomena such as foam formation, turbidity, or particle floating can be observed by visual inspection, supplementing information from other analytical methods 36…”

Section: Methods For Particle Analysismentioning

confidence: 99%

Particles in Therapeutic Protein Formulations, Part 1: Overview of Analytical Methods

Zölls¹,

Tantipolphan²,

Wiggenhorn³

et al. 2012

Journal of Pharmaceutical Sciences

201

159

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Section: Methods For Particle Analysismentioning

confidence: 99%

Particles in Therapeutic Protein Formulations, Part 1: Overview of Analytical Methods

Zölls¹,

Tantipolphan²,

Wiggenhorn³

et al. 2012

Journal of Pharmaceutical Sciences

201

159

View full text Add to dashboard Cite

“…This can be destabilizing for liquid formulations because proteins are exposed to different interfaces during the development process. Furthermore, fill volumes less than the total vial capacity introduce air-water interface which can lead to protein aggregation upon mechanical agitation (39). To study the effect of polyols on the physical stability of proteins under mechanical stress, solutions of mAb-U were shaken at 200 rpm for 5 days at 25±0.1°C in different polyols.…”

Section: Effect Of Polyols On the Physical Stability Of Mab-u Under Mmentioning

confidence: 99%

Opposite Effects of Polyols on Antibody Aggregation: Thermal Versus Mechanical Stresses

Abbas

Sharma²,

Patapoff³

et al. 2011

Pharm Res

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Preferentially excluded polyols increase the conformational stability of proteins but also increase their chemical potential in the solution phase. This increase in free energy can promote precipitation and interfacial adsorption of a protein as these reactions result in a decrease in its free energy. Therefore, addition of polyols can be destabilizing for the physical stability of aqueous protein formulations.

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“…development. [12][13][14]30,31 Agitation of protein solutions can cause formation of insoluble aggregates and particles which contribute to increased turbidity. 14 A mAb mixture (IgG1-A, IgG2-A, and IgG2-B) was used to establish whether mechanisms of agitationinduced aggregation differ from those of temperature-induced aggregation.…”

Section: Agitation-induced Mab Aggregationmentioning

confidence: 99%

“…11 It has also been established that denatured or misfolded mAbs aggregate quickly, 11 often with no detectable accumulation of soluble (oligomeric) species. [12][13][14] Previously, we demonstrated the use of native cation-exchange chromatography (CEX) in solving analytical problems associated with charge heterogeneity, covalent modifications, and dimerization in mAbs. 15,16 Here, we continue to exploit the separation capability of CEX to analyze mixtures of fulllength mAbs and their respective Fab and Fc fragments.…”

Section: Introductionmentioning

confidence: 99%

The use of native cation‐exchange chromatography to study aggregation and phase separation of monoclonal antibodies

et al. 2010

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This study introduces a novel analytical approach for studying aggregation and phase separation of monoclonal antibodies (mAbs). The approach is based on using analytical scale cation-exchange chromatography (CEX) for measuring the loss of soluble monomer in the case of individual and mixed protein solutions. Native CEX outperforms traditional size-exclusion chromatography in separating complex protein mixtures, offering an easy way to assess mAb aggregation propensity. Different IgG1 and IgG2 molecules were tested individually and in mixtures consisting of up to four protein molecules. Antibody aggregation was induced by four different stress factors: high temperature, low pH, addition of fatty acids, and rigorous agitation. The extent of aggregation was determined from the amount of monomeric protein remaining in solution after stress. Consequently, it was possible to address the role of specific mAb regions in antibody aggregation by co-incubating Fab and Fc fragments with their respective full-length molecules. Our results revealed that the relative contribution of Fab and Fc regions in mAb aggregation is strongly dependent on pH and the stress factor applied. In addition, the CEX-based approach was used to study reversible protein precipitation due to phase separation, which demonstrated its use for a broader range of protein-protein association phenomena. In all cases, the role of Fab and Fc was clearly dissected, providing important information for engineering more stable mAb-based therapeutics.

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Relevant shaking stress conditions for antibody preformulation development

Cited by 47 publications

References 23 publications

Particles in Therapeutic Protein Formulations, Part 1: Overview of Analytical Methods

Particles in Therapeutic Protein Formulations, Part 1: Overview of Analytical Methods

Opposite Effects of Polyols on Antibody Aggregation: Thermal Versus Mechanical Stresses

The use of native cation‐exchange chromatography to study aggregation and phase separation of monoclonal antibodies

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