Ion exchange method (IEX) for the analysis of charge variants in biotherapeutic drugs
Robert Birdsall, Thomas Wheat, and Weibin Chen
Waters Corporation (Milford, MA, USA)
Application advantages â– Increase productivity through automated analytical techniques â– Robust method development results in consistent and reproducible results for the validation and quantification of biotherapeutic drug charge variants â– Eliminates the need to prepare additional buffers, simplifies method development and improves Method reproducibility
Waters Solutions
UNIFI® Biopharmaceutical Platform Solutions
ACQUITY UPLC® H-Class System
ACQUITY UPLC Variable Wavelength Ultraviolet (TUV) Detector
Protein-Pak TM Hi Res SP SCX Column
UNIFI Scientific Information System
Key words
Auto • Blend Plus TM technology, cation exchange, an antibody, IEX, SCX, chromatography, biological separation, proteins, methods development, stability
About <br> when mass properties characterization and monitoring of therapeutic proteins, charge variant analysis is critical. Protein modification such as deamidation, N-terminal pyroglutamic acid group modification, isomerization, sialylated glycan, and C-terminal lysine tailoring can lead to the formation of charge variants 1 . In some cases, these changes can affect the binding of therapeutic proteins, biological activity, patient safety, and shelf life.
The biopharmaceutical industry characterizes charge variants by means of ion exchange chromatography (IEX) and isoelectric focusing (IEF) gel electrophoresis. Ion exchange chromatography is particularly useful for biotherapeutic drug development because of its ease of use, wide applicability, and high resolution.
If the charge heterogeneity of therapeutic proteins in biopharmaceutical development is to be characterized in depth, a stable and efficient IEX method is needed. When developing methods, a comprehensive assessment of all possible experimental parameters, such as buffer/ion strength, buffer pH, salt gradient, flow rate, and column temperature, is typically performed. However, in order to systematically evaluate the effect of individual experimental parameters on separation performance, a lengthy iterative process is often required, which involves the preparation and testing of various different component buffers.
Differences in buffer preparation can result in inconsistent results, which in turn can lead to method development time. Waters® Auto•Blend Plus technology utilizes the quaternary solvent management technology of the ACQUITY UPLC H-Class system to solve these problems with pure solvents and high concentration modifier stock solutions. Use Auto•Blend Plus technology to calculate the percentage of each stock solution to achieve the desired pH, reducing errors, reducing consumables and reducing development time.
The UNIFI biopharmaceutical platform solution integrates these features and is ideal for stable method development and easy automation to increase productivity. This application note is intended to demonstrate the high efficiency and stability of the Auto•Blend Plus technology in the optimization of charge variant separation methods using the ion exchange (IEX) method. This application note uses the chimeric monoclonal antibody infliximab as an example of a therapeutic protein application.
To download the application note, please click: http://?cid=511436&lid=134776834&locale=zh_CN
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