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Self-packed core shell nano liquid chromatography columns and silica-based monolithic trap columns for targeted
Henriette Sjaanes Berg1, Kristina Erikstad Seterdal1, Tone Smetop1
1Department of Chemistry, University of Oslo, Post Box 1033, Blindern, NO-0315 Oslo, Norway.
Journal of Chromatography. A
|April 8, 2017
Summary
Researchers developed cost-effective, self-prepared nano liquid chromatography (nLC) columns for targeted proteomics. These in-house columns offer comparable performance to commercial options and enable sensitive detection of cancer biomarkers.
Area of Science:
- Analytical Chemistry
- Proteomics
- Biochemistry
Background:
- Self-preparation of nano liquid chromatography (nLC) columns offers significant cost and flexibility advantages.
- Targeted proteomics requires high-performance separation techniques for accurate biomarker quantification.
- Developing robust and reproducible in-house nLC columns is crucial for advancing proteomic research.
Purpose of the Study:
- To evaluate and optimize methods for self-preparation of particle-packed nLC columns and fritless monolithic trap columns.
- To assess the performance of in-house prepared columns in terms of peak capacity and retention time repeatability.
- To demonstrate the application of these self-prepared columns for targeted biomarker detection in cancer cells.
Main Methods:
- Particle-packed nLC columns were prepared using Accucore core-shell particles (C18) with magnetic stirring and pressure-driven filling.
- Frittless silica-based monolithic trap columns were manufactured using a reproducible recipe.
- Column performance was evaluated by comparing peak capacities and retention time repeatability with commercial columns.
- The prepared columns were utilized for the targeted detection of the enzyme CYP27A1 in cancer cell samples.
Main Results:
- Self-prepared nLC columns demonstrated comparable peak capacities to commercial columns and satisfactory retention time repeatability.
- In-house packed columns using fused silica capillaries matched the performance of those using commercial capillary housings.
- Monolithic trap columns (C8) significantly reduced peak widths compared to standard C18 trap columns.
- The developed in-house columns successfully enabled targeted detection of CYP27A1 in cancer cells.
Conclusions:
- Self-preparation of nLC and trap columns provides a cost-effective and flexible alternative to commercial options.
- In-house prepared columns meet the performance requirements for targeted proteomics applications.
- The developed monolithic trap columns offer enhanced separation efficiency.
- These readily prepared columns facilitate the sensitive detection of clinically relevant biomarkers like CYP27A1 in cancer research.