Capillary ion-exchange chromatography with nanogram sensitivity for the analysis of monoclonal antibodies

Rea, Jennifer C.; Freistadt, Benny S.; McDonald, D. G.; Farnan, Dell; Wang, Yajun Jennifer

doi:10.1016/j.chroma.2015.11.002

Cited by 11 publications

(8 citation statements)

References 25 publications

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“…Conventional ion-exchange chromatography needs to be elaborated for every single mAb, is a time-consuming procedure, and needs several micrograms of mAb for each injection. This led to the development of highly sensitive alternatives like capillary ion-exchange chromatography (Rea et al 2015 ) and high-performance ion-exchange chromatography (Kishishita et al 2015 ) to analyze acidic, main, and basic mAb fractions. To handle multiple samples in a short time and evaluate different process strategies, Wheeler et al ( 2014 ) developed a microchip zone electrophoresis for high throughput analysis of mAbs.…”

Section: The Factor “Product Quality” In Mab Productionmentioning

confidence: 99%

Advances in recombinant antibody manufacturing

Kunert

Reinhart

2016

Appl Microbiol Biotechnol

346

228

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Since the first use of Chinese hamster ovary (CHO) cells for recombinant protein expression, production processes have steadily improved through numerous advances. In this review, we have highlighted several key milestones that have contributed to the success of CHO cells from the beginning of their use for monoclonal antibody (mAb) expression until today. The main factors influencing the yield of a production process are the time to accumulate a desired amount of biomass, the process duration, and the specific productivity. By comparing maximum cell densities and specific growth rates of various expression systems, we have emphasized the limiting parameters of different cellular systems and comprehensively described scientific approaches and techniques to improve host cell lines. Besides the quantitative evaluation of current systems, the quality-determining properties of a host cell line, namely post-translational modifications, were analyzed and compared to naturally occurring polyclonal immunoglobulin fractions from human plasma. In summary, numerous different expression systems for mAbs are available and also under scientific investigation. However, CHO cells are the most frequently investigated cell lines and remain the workhorse for mAb production until today.

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Section: The Factor “Product Quality” In Mab Productionmentioning

confidence: 99%

Advances in recombinant antibody manufacturing

Kunert

Reinhart

2016

Appl Microbiol Biotechnol

346

228

View full text Add to dashboard Cite

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“…The majority of applications are performed using 4.6 or 2.1 mm id column formats. Rea and Farnan reported on the use of capillary columns formats, i.e., 400 μm id columns packed with 5 μm pellicular strong cation‐exchange particles and 300 μm id columns packed with 1.7 μm nonporous weak cation exchange particles for the separation of mAb charge variants .…”

Section: Native Lc Modesmentioning

confidence: 99%

Advances in native high‐performance liquid chromatography and intact mass spectrometry for the characterization of biopharmaceutical products

Tassi

Vos

Chatterjee

et al. 2017

J of Separation Science

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The characterization of biotherapeutics represents a major analytical challenge. This review discusses the current state‐of‐the‐art in analytical technologies to profile biopharma products under native conditions, i.e., the protein three dimensional conformation is maintained during liquid chromatographic analysis. Native liquid‐chromatographic modes that are discussed include aqueous size‐exclusion chromatography, hydrophobic interaction chromatography, and ion‐exchange chromatography. Infusion conditions and the possibilities and limitations to hyphenate native liquid chromatography to mass spectrometry are discussed. Furthermore, the applicability of native liquid‐chromatography methods and intact mass spectrometry analysis for the characterization of monoclonal antibodies and antibody–drug conjugates is discussed.

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“…Routine monitoring of mAb charge heterogeneity is an integral part of any product quality and lifecycle management plan. Charge sensitive analytical methods commonly employed for mAb charge heterogeneity characterization and monitoring include capillary isoelectric focusing (CIEF) [ 16 , 17 ], imaged capillary isoelectric focusing (ICIEF) [ 18 ], capillary zone electrophoresis (CZE) [ 19 – 21 ], ion exchange chromatography (IEX) [ 22 – 24 ], and free-flow electrophoresis (FFE) [ 25 ]. Capillary and microchip-based techniques [ 26 ] afford very high separation efficiency even for very rapid separations (<15 min), while consuming very little sample.…”

Section: Introductionmentioning

confidence: 99%

Qualification of NISTmAb charge heterogeneity control assays

Turner

Schiel

2018

Anal Bioanal Chem

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The NISTmAb is a monoclonal antibody Reference Material from the National Institute of Standards and Technology; it is a class-representative IgG1κ intended serve as a pre-competitive platform for harmonization and technology development in the biopharmaceutical industry. The publication series of which this paper is a part describes NIST’s overall control strategy to ensure NISTmAb quality and availability over its lifecycle. In this paper, the development and qualification of methods for monitoring NISTmAb charge heterogeneity are described. Capillary zone electrophoresis (CZE) and capillary isoelectric focusing (CIEF) assays were optimized and evaluated as candidate assays for NISTmAb quality control. CIEF was found to be suitable as a structural characterization assay yielding information on the apparent pI of the NISTmAb. CZE was found to be better suited for routine monitoring of NISTmAb charge heterogeneity and was qualified for this purpose. This paper is intended to provide relevant details of NIST’s charge heterogeneity control strategy to facilitate implementation of the NISTmAb as a test molecule in the end user’s laboratory. Graphical AbstractRepresentative capillary zone electropherogram of the NIST monoclonal antibody (NISTmAb). The NISTmAb is a publicly available research tool intended to facilitate advancement of biopharmaceutical analytics. Electronic supplementary materialThe online version of this article (10.1007/s00216-017-0816-6) contains supplementary material, which is available to authorized users.

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Capillary ion-exchange chromatography with nanogram sensitivity for the analysis of monoclonal antibodies

Cited by 11 publications

References 25 publications

Advances in recombinant antibody manufacturing

Advances in recombinant antibody manufacturing

Advances in native high‐performance liquid chromatography and intact mass spectrometry for the characterization of biopharmaceutical products

Qualification of NISTmAb charge heterogeneity control assays

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