Development of an effective sample preparation method for the proteome analysis of body fluids using 2-D gel electrophoresis.

Author(s): Joo WA, Lee DY, Kim CW

Publication: Biosci Biotechnol Biochem, 2003, Vol. 67, Page 1574

PubMed ID: 12913303 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to optimize protein extraction from human body fluids using a combined centrifugal filter device and a sample treating buffer for analysis with 2-D gel electrophoresis. They compared this method with traditional methods of protein extraction for plasma, amniotic fluid, urine, and tears.

   Summary of Findings:

   The authors report that extraction using a centrifugal filter device and the zwitterionic detergent CHAPS resulted in a nearly 90% protein recovery and high quality 2-D gel separation. Biospecimen location specific results are as follows: Plasma: Low-molecular-weight gel spots were more apparent with proteins extracted using the centrifugal-filter/CHAPS method than the hot sodium dodecyl sulfate (SDS) extraction. Amniotic fluid: Hot SDS and centrifugal-filter/CHAPS extractions yielded comparable results. Urine: Low-molecular-weight gel spots were significantly increased for proteins extracted using the centrifugal-filter/CHAPS methods compared to those extracted with dialysis alone. Tears: Many protein spots were apparent for proteins extracted with the centrifugal-filter/CHAPS method, which enabled good isoelectric focusing.

   Biospecimens

   • Bodily Fluid - Plasma

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Protein	1D/2D gels

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	Centrifugal filter device/CHAPS Hot SDS Dialysis

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