Thesis
The development and evaluation of a method for the quantitative determination of a monoclonal antibody using capillary electrophoresis-high resolution mass spectrometry
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2020
- Thesis identifier
- T16434
- Person Identifier (Local)
- 201777787
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- Biotherapeutics are becoming of greater importance within the pharmaceutical industry, with companies expanding their portfolios into this area and an increasing number of biotherapeutic regulatory submissions and approvals. As a result of this move away from traditional small molecules progresses the methods used to support the data for the submissions have to evolve and new techniques explored to ensure the quality of data and meet the analytical challenges that this shift in strategy presents. LC-MS has been the main technique involved in the production of toxicokinetic (TK) and pharmacokinetic (PK) data for a number of years and has been reliable for quantification of small molecules. LC-MS using a ‘bottom-up’ approached based on enzyme digestion and surrogates’ peptides along with plate-based techniques such as enzyme linked immunosorbent assay (ELISA) have been used for quantification of biotherapeutics however these techniques both have restrictions and limitations. This work looked to develop a method that could fill these gaps for the quantification of monoclonal antibodies (mAbs) using capillary electrophoresis-high resolution mass spectrometry (CE-HRMS). The final method that was developed for quantification was based on reduction to the light and heavy chains and used a capillary electrophoresis (CE) separation with a discontinuous buffer system and two stage application of pressure to achieve a method that produces good peak shape; analyte response (in terms of peak intensity) and reasonable run times. The method used electrokinetic injection as a means of introducing the sample to the capillary. The data are greatly improved by the used of an internal standard to counteract the variation that is introduced as an artefact of the electrokinetic injection. The CE-MS method had a low limit of quantification for the mAb of 50 ng/mL in human plasma and achieved partial resolution of the mAb light and heavy chains in a run time of 15 minutes. This work shows that CE-MS has the potential to be used to produce PK and TK data based on a ‘middle-up’ approach, that is more representative of the circulating levels of the intact unmodified mAb
- Resource Type
- Note
- Previously held under moratorium in Chemistry department (GSK) from 17 June 2020 until 17 June 2022.
- DOI
- Funder
- Embargo Note
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