Centrifugal ultrafiltration as tool for low abundance proteins isolation

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Article Title:	Centrifugal ultrafiltration as tool for low abundance proteins isolation
Authors:	Ioana Lancrajan, Doru Ardelean, Svetlana Kogalniceanu
Affiliation:	„Vasile Goldiş” Western University of Arad, Faculty of Life Sciences, Romania
Abstract:	Because of the large amount of albumin and others “high abundance proteins” (HAP), the “dynamic range” and heterogeneity of proteins, the separation, isolation, characterization and quantifying of “low abundance proteins” LAP from serum, most of them important diagnostically marchers, is technically difficult. The aim this study is to test a simple and cheap method to reduce the serum protein complexity in order to identify and quantify LAP, particularly growth hormone. Ultrafiltration was used as preparative method to remove HAP from serum and enrich the sample in LAP. Denaturation seems to be essential before centrifugal ultrafiltration (Georgiou, 2001/ Tirumalai, 2003/ Harper, 2004). Filtration was performed under denaturing conditions and recovery of hGH calculated. The higher recovery rate (46%) was attempted when the samples was denatured before centrifugal ultrafiltration in buffer containing RapiGest, compared with denaturation in SDS (6,2% recovery). The results are helpful for procedures of quantitative determination of LAP e.g. hGH from serum.
Keywords:	ultrafiltration, denaturation, recovery, low abundance protein, quantify
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*Correspondence:	Ioana Lancrajan, „Vasile Goldiş” Western University of Arad, Faculty of Life Sciences, Rebreanu st. 91-93; email: Lancrajan _ioana@yahoo.com