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Updated: Jul 17, 2026
Preparation of Mitochondria from Ovarian Cancer Tissues and Control Ovarian Tissues for Quantitative Proteomics Analysis
Published on: November 18, 2019
Membrane proteome analysis of microdissected ovarian tumor tissues using capillary isoelectric
Weijie Wang1, Tong Guo, Paul A Rudnick
1Calibrant Biosystems, 910 Clopper Road, Suite 220N, Gaithersburg, Maryland 20878, USA.
Abstract:
This work expands our tissue proteome capabilities from the analysis of soluble proteins in previous studies to the examination of membrane proteins within the pellets of enriched and selectively isolated tumor cells procured from microdissected tissue specimens. The pellets of targeted ovarian tumor cells are treated by two different membrane protein extraction methods, including the use of detergent and organic solvent. The detergent-based membrane protein preparation protocol not only extracts proteins effectively from cell pellets but also is compatible with subsequent proteome analysis using combined capillary isoelctric focusing/nano reversed-phase liquid chromatography separations coupled with nano electrospray ionization mass spectrometry. Among proteins identified from an amount of pellet equivalent to 20 000 cells, 773 proteins are predicted to contain one or more transmembrane domains, corresponding to 22% membrane proteome coverage within the SwissProt Human protein sequence entries.
Insights
This study advances tissue proteome analysis by examining membrane proteins in isolated ovarian tumor cells. A novel detergent-based method effectively extracts these proteins for mass spectrometry, identifying 773 transmembrane proteins.
Area of Science:
- Biochemistry
- Proteomics
- Oncology
Background:
- Previous tissue proteome studies focused on soluble proteins.
- Analysis of membrane proteins in isolated tumor cells presents unique challenges.
Purpose of the Study:
- To expand tissue proteome analysis to include membrane proteins from isolated tumor cells.
- To develop and validate an effective membrane protein extraction method for proteomic analysis.
Main Methods:
- Selective isolation of tumor cells from microdissected tissue specimens.
- Application of two membrane protein extraction methods: detergent-based and organic solvent-based.
- Proteomic analysis using capillary isoelectric focusing/nano reversed-phase liquid chromatography and nano electrospray ionization mass spectrometry.
Main Results:
- The detergent-based protocol effectively extracted membrane proteins and was compatible with downstream proteomic analysis.
- Analysis of 20,000 cells yielded 773 proteins predicted to have transmembrane domains.
- Achieved 22% coverage of the human membrane proteome within SwissProt entries.
Conclusions:
- The developed method enables comprehensive analysis of the tumor cell membrane proteome.
- This advancement is crucial for understanding ovarian cancer biology and identifying potential therapeutic targets.
