Capillary isoelectric focusing with UV fluorescence imaging detection enables direct charge heterogeneity characterization of erythropoietin drug products

Li, Xiang; Yu, Lei; Shi, Xingming; Chun-ming, Rao; Zhou, Yong

doi:10.1016/j.chroma.2021.462043

Cited by 9 publications

(6 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…CIEF and iCIEF have matured into techniques that are routinely applied in analytical laboratories, especially in the biopharmaceutical industry. Exemplary applications include, but are not limited to, monoclonal antibodies [192,[209][210][211][212][213][214][215], bispecific antibodies [216][217][218], ADCs [219,220], PEGylated proteins [221,222], glycoproteins, in particular erythropoietin [213,223,224], and fusion proteins [213,225]. Because this subsection is part of the former "proteins" section, which was extended to all biopharmaceuticals, it should not go unmentioned that the range of applications has been expanded beyond proteins and includes oligosaccharides [226], LNPs and related mRNA vaccines [227][228][229][230], virus-like particles [231,232], poliovirus [230,233], adeno-associated virus [230,[234][235][236][237], papillomavirus [238], and bacteria and yeast cells [230].…”

Section: Cief and Iciefmentioning

confidence: 99%

See 1 more Smart Citation

Strategies for capillary electrophoresis: Method development and validation for pharmaceutical and biological applications—Updated and completely revised edition

et al. 2023

View full text Add to dashboard Cite

This review is in support of the development of selective, precise, fast, and validated capillary electrophoresis (CE) methods. It follows up a similar article from 1998, Wätzig H, Degenhardt M, Kunkel A. “Strategies for capillary electrophoresis: method development and validation for pharmaceutical and biological applications,” pointing out which fundamentals are still valid and at the same time showing the enormous achievements in the last 25 years. The structures of both reviews are widely similar, in order to facilitate their simultaneous use. Focusing on pharmaceutical and biological applications, the successful use of CE is now demonstrated by more than 600 carefully selected references. Many of those are recent reviews; therefore, a significant overview about the field is provided. There are extra sections about sample pretreatment related to CE and microchip CE, and a completely revised section about method development for protein analytes and biomolecules in general. The general strategies for method development are summed up with regard to selectivity, efficiency, precision, analysis time, limit of detection, sample pretreatment requirements, and validation.

show abstract

Section: Cief and Iciefmentioning

confidence: 99%

“…Urea is still one of the most used additives to prevent precipitation [192, 209–211, 216, 219, 223, 224, 267, 268]. However, several other substances have proven their usefulness.…”

Section: Proteins and Biopharmaceuticalsmentioning

confidence: 99%

Strategies for capillary electrophoresis: Method development and validation for pharmaceutical and biological applications—Updated and completely revised edition

et al. 2023

View full text Add to dashboard Cite

show abstract

“…The pretreatment process must be carried out before analyzing recombinant human erythropoietin (rhEPO) drug products (DPs) by general analysis methods because of their complex composition. Nevertheless, an imaged capillary isoelectric focusing‐UV fluorescence imaging detection method could be used for the direct detection of rhEPO in DPs without pretreatment [53]. The applicability of the method was confirmed by evaluating eight different commercial rhEPO‐DPs with a concentration of 2000–10 000 IU/mL.…”

Section: Applicationsmentioning

confidence: 99%

Applications of capillary electrophoresis in the fields of environmental, pharmaceutical, clinical, and food analysis (2019–2021)

Wang

Gong

Liu

et al. 2022

J of Separation Science

View full text Add to dashboard Cite

So far, the potential of capillary electrophoresis in the application fields has been increasingly excavated due to the advantages of simple operation, short analysis time, high‐resolution, less sample consumption, and low cost. This review examines the implementations and advancements of capillary electrophoresis in different application fields (environmental, pharmaceutical, clinical, and food analysis) covering the literature from 2019 to 2021. In addition, ultrasmall sample injection volume (nanoliter range) and short optical path lead to relatively low concentration sensitivity of the most frequently used ultraviolet‐absorption spectrophotometric detection, so the pretreatment technology being developed has been gradually utilized to overcome this problem. Despite the review being focused on the development of capillary electrophoresis in the fields of environmental, pharmaceutical, clinical, and food analysis, the new sample pretreatment techniques of microextraction and enrichment fit excellently to capillary electrophoresis in recent three years are also described briefly.

show abstract

“…In a recent publication, Li et al. reported the use of FL detection to monitor charge variants of low levels of erythropoietin in simulated drug product formulations [37]. Another drawback of CIEF technology is lack of resolution among co‐formulated molecules with similar p I values.…”

Section: Introductionmentioning

confidence: 99%

“…The conventional iCIEF and IEX assays using UV detection lack the needed sensitivity [36] and dynamic range to simultaneously quantify charge variants of multiple components in wide ratios, such as FDC products with ratios ranging from 1:6 to 1:40. In a recent publication, Li et al reported the use of FL detection to monitor charge variants of low levels of erythropoietin in simulated drug product formulations [37]. Another drawback of CIEF technology is lack of resolution among co-formulated molecules with similar pI values.…”

Section: Introductionmentioning

confidence: 99%

Dual‐detection approach for a charge variant analysis of monoclonal antibody combination products using imaged capillary isoelectric focusing

Candreva

Esterman

et al. 2022

Electrophoresis

View full text Add to dashboard Cite

The clinical benefits of treatments with a combination of two or more therapeutic monoclonal antibodies (mAbs) have emerged in recent years. Imaged capillary isoelectric focusing is a frequently used technology in the biopharmaceutical industry for charge variant analysis of protein therapeutics. However, with the wide concentration ranges of combination products, one component may fall within the linear detection range, whereas the other does not. Here, we report a novel methodology to explore charge variants of mAb mixtures using multiple detection techniques simultaneously. We use ultraviolet absorbance to monitor the charge variants of the high-concentration component and native fluorescence (FL) to monitor the variants of the low-concentration one. Charge variants of mixtures that span 40-fold in ratio differences can be accurately quantified with this approach. In contrast to the conventional methods, it is not necessary to prepare and analyze two samples at different concentrations and combine the results for combination product testing. Additionally, the use of FL detection enables the charge variant analysis of highly potent/low abundant mAbs in a mixture. This methodology is more quality-control friendly and efficient for the charge variant analysis of combination products with wide ratios.

show abstract

Capillary isoelectric focusing with UV fluorescence imaging detection enables direct charge heterogeneity characterization of erythropoietin drug products

Cited by 9 publications

References 11 publications

Strategies for capillary electrophoresis: Method development and validation for pharmaceutical and biological applications—Updated and completely revised edition

Strategies for capillary electrophoresis: Method development and validation for pharmaceutical and biological applications—Updated and completely revised edition

Applications of capillary electrophoresis in the fields of environmental, pharmaceutical, clinical, and food analysis (2019–2021)

Dual‐detection approach for a charge variant analysis of monoclonal antibody combination products using imaged capillary isoelectric focusing

Contact Info

Product

Resources

About