Enhancement of covalent aggregate quantification of protein therapeutics by non-reducing capillary gel electrophoresis using sodium hexadecyl sulfate (CE-SHS)

Guan, Qian; Knihtila, Ryan; Atsma, Jennifer; Tulsan, Rekha; Singh, Surinder; Kar, S.; Beckman, Jeff; Ding, James; Li, Zheng Jian

doi:10.1016/j.jchromb.2020.122230

Cited by 10 publications

(7 citation statements)

References 27 publications

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“…During electrophoresis, the SDS on the surface of the LC–SDS complex dissociates, forming a series of LC–SDS complexes with different degrees of negative charges, which leads to tailing. For LC–SHS complexes, stronger hydrophobicity can form more stable LC–SHS complexes through hydrophobic interactions than LC–SDS complex [23], so the dissociation reaction of SHS is slight. However, the reason why LC and SDS cannot be stably combined is still unknown and may be related to its microstructure or special amino acid sequence.…”

Section: Resultsmentioning

confidence: 99%

“…In view of the previous use of STS and SHS to mitigate the HC or intact mAb peak artifacts in CE-SDS [21][22][23], we tried to use CE-SHS and CE-STS to optimize the LC peak shape of mAb-A, that is, replacing the SDS of the sample and gel buffer with SHS and STS, respectively. Both CE-SHS and CE-STS have significantly optimized peak shapes (Figure 5).…”

Section: 3mentioning

confidence: 99%

“…Molina et al [20] reported HMW artifacts caused by incomplete denaturation of mAbs, and they increased the SDS concentration, denaturation temperature, and incubation time to eliminate the artifacts. Guan et al [21][22][23] also reported HMW artifacts in reducing and nonreducing modes. They attributed this phenomenon to the problem of binding between the ionic surfactant SDS and the antibody molecule.…”

Section: Introductionmentioning

confidence: 96%

“…Guan et al. [21–23] also reported HMW artifacts in reducing and nonreducing modes. They attributed this phenomenon to the problem of binding between the ionic surfactant SDS and the antibody molecule.…”

Section: Introductionmentioning

confidence: 99%

“…Finally, the use of the longer alkyl chain anionic surfactants sodium tetradecyl sulfate (STS) and sodium hexadecyl sulfate (SHS) as alternatives to SDS achieved the best optimization effect. SHS and STS have been previously reported for the optimization of HC or intact mAbs [16,22,23], but they have never been applied to improve LC peak performance as reported in this article. Here, we proved that SHS was also suitable for eliminating the tailing peak caused by the limitation of the complexation of SDS and LC in reducing CE-SDS, thereby improving the peak shape and providing a more accurate and stable analysis method.…”

Section: Introductionmentioning

confidence: 99%

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Elimination of light chain tailing in reducing capillary electrophoresis with sodium dodecyl sulfate analysis of a monoclonal antibody

et al. 2022

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Capillary electrophoresis with sodium dodecyl sulfate (CE‐SDS) is a common analytical technique for investigating the purity and molecular size heterogeneity of monoclonal antibody (mAb) drugs. In reducing CE‐SDS analysis of mAb‐A, the light chain (LC) peak exhibited severe tailing, seriously affecting the purity analysis. The purposes of this investigation are to clarify the source of tailing and develop a more appropriate CE‐SDS method to eliminate LC tailing. The degree of LC tailing was closely related to the mAb concentration, SDS concentration, and injection amount, and more hydrophobic detergents, such as sodium hexadecyl sulfate (SHS) and sodium tetradecyl sulfate (STS), could be used instead of SDS to obtain better peak shapes. The results also indicated that the tailing was caused by the binding problem associated with SDS, and SHS/STS could provide a more stable and uniform complexation for the LC. In summary, the method we developed successfully eliminated the LC tailing and provided a robust characterization of mAb‐A in reducing CE‐SDS analysis.

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Section: Resultsmentioning

confidence: 99%

Section: 3mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Elimination of light chain tailing in reducing capillary electrophoresis with sodium dodecyl sulfate analysis of a monoclonal antibody

et al. 2022

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Applications of capillary electrophoresis for biopharmaceutical product characterization

2021

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Capillary electrophoresis (CE), after being introduced several decades ago, has carved out a niche for itself in the field of analytical characterization of biopharmaceutical products. It does not only offer fast separation, high resolution in miniaturized format, but equally importantly represents an orthogonal separation mechanism to high‐performance liquid chromatography. Therefore, it is not surprising that CE‐based methods can be found in all major pharmacopoeias and are recommended for the analysis of biopharmaceutical products during process development, characterization, quality control, and release testing. Different separation formats of CE, such as capillary gel electrophoresis, capillary isoelectric focusing, and capillary zone electrophoresis are widely used for size and charge heterogeneity characterization as well as purity and stability testing of therapeutic proteins. Hyphenation of CE with MS is emerging as a promising bioanalytical tool to assess the primary structure of therapeutic proteins along with any impurities. In this review, we confer the latest developments in capillary electrophoresis, used for the characterization of critical quality attributes of biopharmaceutical products covering the past 6 years (2015–2021). Monoclonal antibodies, due to their significant share in the market, have been given prioritized coverage.

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Recent biopharmaceutical applications of capillary electrophoresis methods on recombinant DNA technology‐based products

et al. 2021

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Recent biopharmaceutical applications of capillary electrophoresis methods on recombinant DNA technology-based productsBiopharmaceuticals (recombinant technology-based products, vaccines, whole blood and blood components, gene therapy, cells, tissues, etc.,) are described as biological medical products produced from various living sources such as human, microbial, animal, and so on by manufacturing, extraction, or semi-synthesis. They are complex molecules having high molecular weights. For their safety and efficacy, their structural, clinical, physicochemical, and chemical features must be carefully controlled, and they must be well characterized by analytical techniques before the approval of the final product. Capillary electrophoresis (CE) having versatile modes can provide valuable safety and efficacy information, such as amino acid sequence, size variants (low and high molecular weight variants), charged variants (acidic and basic impurities), aggregates, N-linked glycosylation, and O-linked glycosylation. There are numerous applications of CE in the literature. In this review, the most significant and recent studies on the analysis of recombinant DNA technology-based products using different CE modes in the last ten years have been overviewed. It was seen that the researches mostly focus on the analysis of mAbs and IgG. In addition, in recent years, researchers have started to prefer CE combined mass spectrometry (MS) techniques to provide a more detailed characterization for protein and peptide fragments.

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Enhancement of covalent aggregate quantification of protein therapeutics by non-reducing capillary gel electrophoresis using sodium hexadecyl sulfate (CE-SHS)

Cited by 10 publications

References 27 publications

Elimination of light chain tailing in reducing capillary electrophoresis with sodium dodecyl sulfate analysis of a monoclonal antibody

Elimination of light chain tailing in reducing capillary electrophoresis with sodium dodecyl sulfate analysis of a monoclonal antibody

Applications of capillary electrophoresis for biopharmaceutical product characterization

Recent biopharmaceutical applications of capillary electrophoresis methods on recombinant DNA technology‐based products

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