Influence of pH, buffer species, and storage temperature on physicochemical stability of a humanized monoclonal antibody LA298

Zheng, Jack; Janis, Linda J.

doi:10.1016/j.ijpharm.2005.10.024

Cited by 107 publications

(69 citation statements)

References 10 publications

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“…Incubating the mAb samples for 15 and 40 hours 5-6 intensive but overlapped peaks of acidic variants could be observed (Figure 4c,d). These results correspond with the statements of others, that is, at high temperature and at high pH the deamidation (the most common covalent modifications for protein pharmaceuticals) becomes faster [3]. A similar increase in acidic variants after stress (0,5 M phosphate, pH 8.0 at 40°C for 8 day) was obtained by Han et al using conventional and microchip CZE [14].…”

Section: Study Of Solution and Thermal Stability Of Rituximabsupporting

confidence: 90%

“…(The dilution of the formulated product MabThera reduces the stabilizer effect of the nonionic surfactant and citrate buffer additives. While the citrate moderates the deamidation by changing the conformation of the protein, the phosphate ions may catalyze the deamidation [3].) Incubating the undiluted MabThera at 55°C for 15 hours it practically did not undergo any transformation (Figure 4e).…”

Section: Study Of Solution and Thermal Stability Of Rituximabmentioning

confidence: 99%

“…While the size heterogeneity of the monoclonal antibodies was studied by size-exclusion chromatography [3,4], peptide mapping [4], LC-MS [5] or gel electrophoresis [6][7][8], the charge heterogeneity was mostly studied by cation-exchange chromatography [3,4]. The different methods of capillary electrophoresis (CZE, CGE, CIEF) are useful in the complex analysis of the mAbs.…”

Section: Introductionmentioning

confidence: 99%

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Analysis of Rituximab, A Therapeutic Monoclonal Antibody by Capillary Zone Electrophoresis

G¹

2014

J Chromatogr Sep Tech

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This paper focuses on the applicability of capillary zone electrophoresis (CZE) using uncoated fused silica capillaries for the determination of heterogeneity of rituximab (MabThera, Roche) and for the study of its solution and thermal stability. The best resolution for the charge variants of the main component was obtained with a buffer electrolyte containing 800 mM 6-amino caproic acid, 2 mM triethylene tetramine and 0.05% hydroxypropyl methylcellulose at pH = 5.2. It was found that the pH and the components of the buffer used for the electrophoretic separation and for the dilution of the sample prior to the analysis are important to the stability of the rituximab. We demonstrated the rituximab is stable in the pharmaceutical product MabThera due to the stabilizing additives, but the dilution of the MabThera caused a slow formation of acidic variants, while the amount of the basic variants did not change. After incubation of the diluted rituximab at higher temperature several charge variants could be determined by CZE.

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Section: Study Of Solution and Thermal Stability Of Rituximabsupporting

confidence: 90%

Section: Study Of Solution and Thermal Stability Of Rituximabmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Analysis of Rituximab, A Therapeutic Monoclonal Antibody by Capillary Zone Electrophoresis

G¹

2014

J Chromatogr Sep Tech

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“…[8] Even though deamidation can happen at virtually every stage from the production of an antibody in the cell to storage of the final product, the rate at which this modification tends to occur is highly dependent on the external conditions including buffer composition, pH, and temperature. [70] For example, it has been reported that incubation of IgG samples at basic pH and elevated temperatures drastically increases the likelihood of deamidation reactions occurring in the sample. [8] As these conditions are often part of the typical sample preparation procedure for LC-MS analysis it can be difficult to identify variants created by this type of modification due to the presence of artifacts resulting from sample preparation.…”

Section: Asparagine and Glutamine Deamidationmentioning

confidence: 99%

Microheterogeneity of Recombinant Antibodies: Analytics and Functional Impact

Beyer

Schuster

Jungbauer

et al. 2017

Biotechnology Journal

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Antibodies are typical examples of biopharmaceuticals which are composed of numerous, almost infinite numbers of potential molecular entities called variants or isoforms, which constitute the microheterogeneity of these molecules. These variants are generated during biosynthesis by so‐called posttranslational modification, during purification or upon storage. The variants differ in biological properties such as pharmacodynamic properties, for example, Antibody Dependent Cellular Cytotoxicity, complement activation, and pharmacokinetic properties, for example, serum half‐life and safety. Recent progress in analytical technologies such as various modes of liquid chromatography and mass spectrometry has helped to elucidate the structure of a lot of these variants and their biological properties. In this review the most important modifications (glycosylation, terminal modifications, amino acid side chain modifications, glycation, disulfide bond variants and aggregation) are reviewed and an attempt is made to give an overview on the biological properties, for which the reports are often contradictory. Even though there is a deep understanding of cellular and molecular mechanism of antibody modification and their consequences, the clinical proof of the effects observed in vitro and in vivo is still not fully rendered. For some modifications such as core‐fucosylation of the N‐glycan and aggregation the effects are clear and should be monitored, but with others such as C‐terminal lysine clipping the reports are contradictory. As a consequence it seems too early to tell if any modification can be safely ignored.

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“…As a result, the surface accessibility of mAb-A to acidic pH 4.5 solution was increased, which might cause more fragmentation. It has been reported that a humanized mAb is the most stable in citrate buffer at pH 5.0-5.5 (16). Another study indicates that two mAbs are less stable in citrate buffer, and aggregation is minimal in Tris-HCl buffer at pH 7.2 and 8 (17).…”

Section: Discussion Aggregation Behaviors Influenced By Buffer Speciesmentioning

confidence: 97%

Investigating the Degradation Behaviors of a Therapeutic Monoclonal Antibody Associated with pH and Buffer Species

et al. 2015

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Abstract. This study aimed in understanding the degradation behaviors of an IgG 1 subtype therapeutic monoclonal antibody A (mAb-A) associated with pH and buffer species. The information obtained in this study can augment conventional, stability-based screening paradigms by providing the direction necessary for efficient experimental design. Differential scanning calorimetry (DSC) was used for studying conformational stability. Dynamic light scattering (DLS) was utilized to generate B 22 *, a modified second virial coefficient for the character of protein-protein interaction. Size-exclusion chromatography (SEC) and hydrophobic interaction chromatography (HIC) were employed to separate degradation products. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used for determining the molecular size and liquid chromatography mass spectrometry (LC-MS) were used for identifying the sequence of the separated fragments. The results showed that both pH and buffer species played the roles in controlling the degradation behaviors of mAb-A, but the pH was more significant. In particular, pH 4.5 induced additional thermal transition peaks occurring at a low temperature compared with pH 6.5. A continual temperature-stress study illustrated that the additional thermal transition peaks related to the least stable structure and a greater fragmentation. Although mAb-A showed the comparable conformational structures and an identical amount of aggregates at time zero between the different types of buffer species at pH 6.5, the aggregation formation rate showed a buffer species-dependent discrepancy over a temperature-stress period. It was found that the levels of aggregations associated with the magnitudes of protein-protein interaction forces.

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Influence of pH, buffer species, and storage temperature on physicochemical stability of a humanized monoclonal antibody LA298

Cited by 107 publications

References 10 publications

Analysis of Rituximab, A Therapeutic Monoclonal Antibody by Capillary Zone Electrophoresis

Analysis of Rituximab, A Therapeutic Monoclonal Antibody by Capillary Zone Electrophoresis

Microheterogeneity of Recombinant Antibodies: Analytics and Functional Impact

Investigating the Degradation Behaviors of a Therapeutic Monoclonal Antibody Associated with pH and Buffer Species

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