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A cost-effective method to get insight into the peritoneal dialysate effluent proteome
J E Araújo1, S Jorge1, F Teixeira E Costa2
1UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal; Bioscope Research Group, UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal; ProteoMass Scientific Society, Madan Park, Rua dos Inventores, 2825-152 Caparica, Portugal.
Two cost-effective proteomics methods using acetonitrile and dithiothreitol successfully analyze peritoneal dialysate effluent. Dithiothreitol shows promise for identifying biomarkers of peritoneal degradation in dialysis patients.
Area of Science:
- Proteomics
- Biochemistry
- Nephrology
Background:
- Renal failure is a growing global health concern, with peritoneal dialysis (PD) offering improved patient quality of life.
- Peritoneal membrane permeability loss limits PD duration, necessitating biomarkers for degradation.
- Identifying peritoneal dialysate effluent (PDE) proteome changes is crucial for PD advancement.
Purpose of the Study:
- To assess acetonitrile (ACN) and dithiothreitol (DTT) as cost-effective proteomics tools for PDE analysis.
- To identify proteins and biological pathways affected in patients undergoing PD.
- To evaluate the potential of these methods for discovering biomarkers of peritoneal degradation.
Main Methods:
- Protein depletion using acetonitrile (ACN) and protein equalization using dithiothreitol (DTT) on PDE samples.
- Two-dimensional gel electrophoresis (2D-PAGE) and peptide mass fingerprinting (PMF) for protein identification.
- Gene Ontology (GO) term annotation to analyze affected biological pathways.
Main Results:
- ACN depleted high-abundance proteins like albumin, enriching apolipoproteins.
- DTT equalized protein levels, reducing albumin and enriching immunoglobulins.
- 72 unique proteins were identified, with extracellular proteins in regulatory binding pathways being most affected.
Conclusions:
- ACN and DTT are effective, economical, and minimal-reagent proteomics tools for PDE analysis.
- Both methods identified affected biological pathways in PD patients, primarily extracellular proteins involved in regulation.
- DTT is a promising method for biomarker discovery due to its equalizing effect, allowing interrogation of more proteins.
