Charge Heterogeneity Analysis

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Charge heterogeneity is often evaluated during antibody drug development, as it is a universal feature of antibody therapeutic products and a regulatory requirement. Charge heterogeneity analysis helps to understand key attributes that may affect the therapeutic stability of an antibody and is important to minimize risk and compress drug development timelines. Creative Proteomics utilizes a range of advanced technologies to perform comprehensive charge variant analysis and testing, allowing for early and informed decisions on product exploitability.

What is Charge Heterogeneity Analysis?

Antibody therapeutic products often undergo aggregation, degradation and various post-translational modifications during production, storage and transportation, resulting in charge inhomogeneities and their corresponding isomers. When the number or structure of charge groups is changed, it will cause heterogeneity of antibodies in charge distribution, which in turn affects the biological activity, pharmacokinetics, immunogenicity and structural stability of antibody drugs, and will have a significant impact on the effectiveness, safety and shelf-life of the drugs in clinical use. Therefore, the characterization and quality control of charge heterogeneity is crucial.

Fig. 1 Trastuzumab charge heterogeneity is highly influenced by asparagine deamidation and aspartic acid isomerization. (Bailey, et al., 2018)

Technology Platform of Charge Heterogeneity Analysis

Charge Heterogeneity Detection Methods

Charge heterogeneity detection is the separation of antibodies based on differences in the charges they carry. Creative Proteomics offers commonly used detection methods including Ion Exchange Chromatography (IEX), Capillary Zone Electrophoresis (CZE), Capillary isoelectric focusing (CIEF), and Imaged capillary isoelectric focusing (iCIEF). Our experienced testing team is well versed in the characteristics of each analytical method and can select the most appropriate method for your demands.

Methods	Description
Ion Exchange Chromatography (IEX)	Based on the difference in surface charge of the antibody isomers in contact with the chromatographic stationary phase, the separation is achieved using a salt concentration gradient, a pH gradient, or a combination of both. High resolution and a versatile tool for charge heterogeneity detection in various samples.
Capillary Zone Electrophoresis (CZE)	Based on the principles of electrophoresis, where charged molecules are separated according to their mobility in an electric field. High resolution separation with excellent reproducibility and sensitivity. It requires only small sample volumes and can handle complex mixtures.
Capillary isoelectric focusing (CIEF)	Based on the difference in Isoelectric point (pI), which reflects the homogeneity of the drug charge and spatial conformation of the protein. High resolution and sensitivity, allowing the detection and quantification of minor charge variants. It also requires small sample volumes.
Imaged capillary isoelectric focusing (iCIEF)	Separating charged molecules based on their isoelectric points (pI). High resolution and sensitivity, allowing for the detection of minor charge variants that can impact the efficacy and quality of antibody drugs.

Charge Heterogeneity Characterization Methods

In order to perform a detailed characterization of the isolated charge isomers, we coupled the isolated isomers with mass spectrometry. Using fully automated on-line mass spectrometry, we can achieve real-time molecular weight detection and identification of charge isomers directly after isolation, and characterize the molecular weight differences of different charge isomers. Our advanced characterization platform greatly improves the efficiency and accuracy of charge heterogeneity mass spectrometry of antibody drugs, thus providing customers with a comprehensive analysis of the charge heterogeneity of samples.

Advantages of Creative Proteomics

High-throughput charge heterogeneity information
Characterization and stability of antibody drug charge variants analyzed at high resolution in less time.
Have confidence in your results with reproducible, accurate charge heterogeneity detection.

Creative Proteomics' charge heterogeneity analysis technology platform is suitable for the comprehensive characterization of charge variant-related quality attributes as well as charge variant analysis throughout the development process. Our powerful characterization solutions help you make fast, confident decisions to drive optimal antibody drug development from the start. Contact us to learn more about our charge heterogeneity analysis service of antibody drugs. We will be happy to assist you.

Reference

Bailey, A. O.; et al. Charge variant native mass spectrometry benefits mass precision and dynamic range of monoclonal antibody intact mass analysis. mAbs. 2018, 10: 1-12.