Different techniques for urinary protein analysis of normal and lung cancer patients.

Author(s): Tantipaiboonwong P, Sinchaikul S, Sriyam S, Phutrakul S, Chen S T

Publication: Proteomics, 2005, Vol. 5, Page 1140

PubMed ID: 15693063 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this paper was to compare the proteomics of urine prepared by gel filtration followed by ultrafiltration or one of four precipitation methods using three analytical platforms: high-performance liquid chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and two-dimensional (2D) gel electrophoresis.

   Summary of Findings:

   Urine purification via ultrafiltration, acetone or ACN/TFA precipitation yielded similar results, while methanol/chloroform/water and TC/acetone precipitation yielded fewer protein peaks, as determined by RP-HPLC. Urine specimens from healthy and lung cancer patients had distinct protein profiles. SDS-PAGE analysis of differentially prepared urine specimens yielded similar results, although diagnosis-specific differences in protein bands were observed. Results of 2-DE analysis were superior with samples purified by ultrafiltration. Urine specimens subjected to sequential gel filtration and ultrafiltration yielded a number of differentially expressed proteins smaller than 30 kDa among healthy and lung cancer patients; nine of these differentially expressed spots were identified via MS analysis.

   Biospecimens

   • Bodily Fluid - Urine

   Preservative Types

   • Frozen

   Diagnoses:

   • Normal
   • Neoplastic - Carcinoma

   Platform:

   Analyte	Technology Platform
   Protein	1D/2D gels
   Protein	LC-MS or LC-MS/MS
   Protein	MALDI-TOF MS
   Protein	HPLC

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	Ultrafiltration Acetone precipitation Acetonitrile/trifluoroacetic acid precipitation Methanol/water/chloroform precipitation Trichloroacetic acid/acetone precipitation
   1D/2D gels Specific	Technology platform	SDS-PAGE 2D gel

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