Charge profiling and stability testing of biosimilar by capillary isoelectric focusing

Cao, Junzi; Sun, Wen; Gong, Feifei; Liu, Wanhui

doi:10.1002/elps.201300471

Cited by 24 publications

(11 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…cIEF method was developed for confident identification of differences in charge-related characteristics of proteins. The same year, Cao et al described a cIEF analysis of trastuzumab biosimilar in terms of charge profiling and stability testing [61]. They realized optimization on focusing time and separation temperature to get a robust method to analyze charge variants.…”

Section: Applicationsmentioning

confidence: 99%

Cutting-edge capillary electrophoresis characterization of monoclonal antibodies and related products

Gahoual

Beck

Leize‐Wagner

et al. 2016

Journal of Chromatography B

View full text Add to dashboard Cite

Out of all categories, monoclonal antibodies (mAbs), biosimilar, antibody-drug conjugates (ADCs) and Fc-fusion proteins attract the most interest due to their strong therapeutic potency and specificity. Because of their intrinsic complexity due to a large number of micro-heterogeneities, there is a crucial need of analytical methods to provide comprehensive in-depth characterization of these molecules. CE presents some obvious benefits as high resolution separation and miniaturized format to be widely applied to the analysis of biopharmaceuticals. CE is an effective method for the separation of proteins at different levels. capillary gel electrophoresis (CGE), capillary isoelectric focusing (cIEF) and capillary zone electrophoresis (CZE) have been particularly relevant for the characterization of size and charge variants of intact and reduced mAbs, while CE-MS appears to be a promising analytical tool to assess the primary structure of mAbs and related products. This review will be dedicated to detail the current and state-of-the-art CE-based methods for the characterization of mAbs and related products.

show abstract

Section: Applicationsmentioning

confidence: 99%

Cutting-edge capillary electrophoresis characterization of monoclonal antibodies and related products

Gahoual

Beck

Leize‐Wagner

et al. 2016

Journal of Chromatography B

View full text Add to dashboard Cite

show abstract

“…Similarly, there is potential application in the comparison of innovator and biosimilar molecules to reveal differences that may be masked by the complexity of isoelectric profile of the intact molecule. 34 It remains to be seen if the ChromiCE method can be incorporated upstream of a mass spectrometry method; however, the potential wealth of information that may result is quite intriguing.…”

Section: Discussionmentioning

confidence: 99%

Rapid quantitative analysis of monoclonal antibody heavy and light chain charge heterogeneity

Vanam

Schneider

Marlow

2015

mAbs

View full text Add to dashboard Cite

To cite this article: Ram P Vanam, Michael A Schneider & Michael S Marlow (2015) Rapid quantitative analysis of monoclonal antibody heavy and light chain charge heterogeneity, mAbs, 7:6, 1118-1127, DOI: 10.1080DOI: 10. /19420862.2015 An alternative method to traditional 2-dimensional gel electrophoresis (2D-PAGE) and its application in characterizing the inherent charge heterogeneity of chromatographically isolated monoclonal antibody heavy and light chains is described. This method, referred to as ChromiCE, utilizes analytical size-exclusion chromatography (SEC), performed under reducing and denaturing conditions, followed by imaged capillary isoelectric focusing (icIEF) of the chromatographically separated heavy and light chains. Under conditions suitable for the subsequent icIEF analysis, the absolute and relative SEC elution volumes of the heavy and light chains were found to be highly pH dependent, a phenomenon that can be exploited in optimizing chromatographic separation. Compared to 2D-PAGE, the ChromiCE method substantially decreases the time and labor needed to complete the analysis, improves reproducibility, and provides fully quantitative assessment of charge heterogeneity. The ChromiCE methodology was applied to a set of diverse monoclonal antibodies to demonstrate suitability for quantitative charge variant analysis of heavy and light chains. A typical application of ChromiCE in extended characterization and stability studies of a purified antibody is shown.

show abstract

“…cIEF method combines the advantage of IEF and CZE as an automatic technique with high resolution and precise calculation of pI value for each component [28,34]. Ahn et al [27] and Alejandro Cifuentes et al [28,29] reported the analysis of rhuEPO isoforms by cIEF.…”

Section: Introductionmentioning

confidence: 99%

Isoforms analysis of recombinant human erythropoietin by polarity‐reversed capillary isoelectric focusing

Ren

Zhang

et al. 2020

Electrophoresis

View full text Add to dashboard Cite

Recombinant human erythropoietin (rhuEPO) has been extensively used as a pharmaceutical product for treating anemia in the clinic. Glycosylation of rhuEPO was crucial for affecting biological activity, immunogenicity, and pharmacokinetics. Because of the heterogeneity of glycan, the structure of rhuEPO was complex with several isoforms. Characterization of isoforms was important for quality control of rhuEPO. Here, an improved cIEF method has been established and validated. A polarity‐reversed focusing step was used by reversing both the polarity of the voltage and the catholyte and anolyte vials. A weak base (100 mM ammonium hydroxide solution) was used as a chemical mobilizer to make the acidic bands mobilize stably to the detection window. Compared with CZE method in European Pharmacopoeia, the numbers of isoforms and their peak area percentage were highly consistent. Better reproducibility and higher resolution have been obtained by the improved cIEF method. Moreover, in improved cIEF method, the isoelectric points (pI) of each isoform can be calculated and used for identification. It was also the first time that the cIEF method was fully validated for rhuEPO analysis according to the International Conference on Harmonization (ICH) guidelines.

show abstract

Charge profiling and stability testing of biosimilar by capillary isoelectric focusing

Cited by 24 publications

References 40 publications

Cutting-edge capillary electrophoresis characterization of monoclonal antibodies and related products

Cutting-edge capillary electrophoresis characterization of monoclonal antibodies and related products

Rapid quantitative analysis of monoclonal antibody heavy and light chain charge heterogeneity

Isoforms analysis of recombinant human erythropoietin by polarity‐reversed capillary isoelectric focusing

Contact Info

Product

Resources

About