Optimization of CE-SDS Method for Antibody Separation Based on Multi-Users Experimental Practices

Han, Mei; Phan, Duke H.; Nightlinger, Nancy S.; Taylor, Lisa D.; Jankhah, S.; Woodruff, Brian; Yates, Zac; Freeman, Sara M.; Guo, Amy; Balland, Alain; Pettit, Dean K.

doi:10.1365/s10337-006-0825-7

Cited by 25 publications

(18 citation statements)

References 10 publications

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“…Capillary electrophoresis sodium dodecyl sulfate (CE-SDS) has evolved over the last two decades to become the method of choice for release and stability analysis of monoclonal antibody (mAb) therapeutics [1][2][3][4][5][6][7]. The method separates molecule specific fragments and other impurities based on their electrophoretic mobility under denaturing conditions [8].…”

Section: Introductionmentioning

confidence: 99%

Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods

Esterman

Katiyar

Krishnamurthy

2016

Journal of Pharmaceutical and Biomedical Analysis

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Section: Introductionmentioning

confidence: 99%

Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods

Esterman

Katiyar

Krishnamurthy

2016

Journal of Pharmaceutical and Biomedical Analysis

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“…Several disadvantages of SDS-PAGE, including the use of toxic reagents, high intra-and inter-gel effective mobility variability, staining variability, limited quantitative abilities, and its labor-intensive nature, have led to its replacement by capillary-based methodology, capillary gel electrophoresis (CGE) [5][6][7][8][9]. Advantages of performing size analysis via capillary technology include automation, enhanced precision, high-speed analysis, an improved resolution for closely migrating species, and on-line quantitative detection by UV (Coomassie Blue stain sensitivity) [5,8,[10][11][12][13][14] or for higher sensitivity LIF detection (silver stain sensitivity) [8,12,15].…”

Section: Introductionmentioning

confidence: 99%

Development, validation, and implementation of capillary gel electrophoresis as a replacement for SDS‐PAGE for purity analysis of IgG2 mAbs

Lacher

Roberts

et al. 2010

J of Separation Science

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Capillary gel electrophoresis (CGE) methods with UV detection were developed for reduced and non-reduced mAb analysis. These methods can be used to evaluate mAb purity, offering more reproducible quantitation compared with that of traditional SDS-PAGE methods. These CGE methods have been utilized as platform technology for bioprocess development, formulation development, mAb characterization, drug substance/drug product release testing as well as a required methodology for stability testing. We have found these CGE methods to be applicable across a platform of mAbs in preclinical and clinical development, with the majority of mAbs requiring no modification to the method conditions. This methodology has been ICH validated and transferred to several supporting organizations. The data presented herein describes the development of CGE methodology, platform application to mAb purity analysis, ICH validation, reliability metrics, and considerations on technology enhancement for improved performance and throughput.

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“…Owing to certain intrinsic limitations of the SDS PAGE assay, many biopharmaceutical laboratories have shifted efforts into developing CE SDS gel [1,2] assays. CE SDS gel assays have been successfully applied to many therapeutic proteins including monoclonal antibody (mAb) [3][4][5] and protein-based vaccines [6,7], and it is now routinely used in the quality-control environment to assess purity and integrity including the assay validation requirements that are easily achieved [1,8].…”

Section: Introductionmentioning

confidence: 99%

Applications of CE SDS gel in development of biopharmaceutical antibody‐based products

2008

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CE SDS gel technique offers many advantages over the traditional labor-intensive SDS PAGE slab gel technology. The CE-based method has increasingly been applied to many protein analysis applications. Specific examples are provided for monoclonal antibody (mAb), though the technique can be adapted to many other therapeutic protein products. Applications of CE SDS gel method using Beckman PA800 with UV detection are presented and discussed with respect to mAb analysis, such as purity, quantitation of non-glycosylated heavy chain (NGHC) peak, identity, and stability. The stability of mAb is evaluated with respect to formulation buffer, accelerated temperature stress, UV light-exposure, and high pH conditions. Both reducing and non-reducing CE SDS gel conditions were applied and optimized to characterize mAb products. The data presented provides a "taste" of what CE SDS gel method can do to support the development of mAb products from early clone screening for product quality to the final product characterization. Since the CE SDS gel method is automatable, quantitative, robust, and allows for relatively high throughput, it provides both great analytical capacity and product coverage for a wide spectrum of protein product development in biopharmaceutical industry.

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Optimization of CE-SDS Method for Antibody Separation Based on Multi-Users Experimental Practices

Cited by 25 publications

References 10 publications

Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods

Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods

Development, validation, and implementation of capillary gel electrophoresis as a replacement for SDS‐PAGE for purity analysis of IgG2 mAbs

Applications of CE SDS gel in development of biopharmaceutical antibody‐based products

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