AI Article Synopsis
- A new method combines size exclusion chromatography (SEC) and mixed-mode liquid chromatography (LC) for analyzing various components in biopharmaceuticals.
- This approach can measure the active pharmaceutical ingredient (protein) along with different excipients like cations, anions, and sugars, using dual SEC columns for separation and detection through UV and evaporative light scattering detectors.
- The technique has been successfully applied to quantify proteins and excipients in various biopharmaceutical products, including monoclonal antibodies and vaccines, and allows for advanced online characterization with mass spectrometry.
Article Abstract
A methodology based on on-line coupling of size exclusion chromatography (SEC) with mixed-mode liquid chromatography (LC) has been developed. The method allows for simultaneous measurement of a wide range of components in biopharmaceutical drug products. These components include the active pharmaceutical ingredient (protein) and various kinds of excipients such as cations, anions, nonionic hydrophobic surfactant and hydrophilic sugars. Dual short SEC columns are used to separate small molecule excipients from large protein molecules. The separated protein is quantified using a UV detector at 280 nm. The isolated excipients are switched, online, to the Trinity P1 mixed-mode column for separation, and detected by an evaporative light scattering detector (ELSD). Using a stationary phase with 1.7 μm particles in SEC allows for the use of volatile buffers for both SEC and mix-mode separation. This facilitates the detection of different excipients by ELSD and provides potential for online characterization of the protein with mass spectrometry (MS). The method has been applied to quantitate protein and excipients in different biopharmaceutical drug products including monoclonal antibodies (mAb), antibody drug conjugates (ADC) and vaccines.
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| http://dx.doi.org/10.1016/j.chroma.2012.09.012 | DOI Listing |
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