A Comparative Proteomic Study of Human Skin Suction Blister Fluid from Healthy Individuals Using Immunodepletion and iTRAQ Labeling
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- 作者：Andr茅 C. M眉ller ; Florian P. Breitwieser ; Heinz Fischer ; Christopher Schuster ; Oliver Brandt ; Jacques Colinge ; Giulio Superti-Furga ; Georg Stingl ; Adelheid Elbe-B眉rger ; Keiryn L. Bennett
- 刊名：Journal of Proteome Research
- 出版年：2012
- 出版时间：July 6, 2012
- 年：2012
- 卷：11
- 期：7
- 页码：3715-3727
- 全文大小：644K
- 年卷期：v.11,no.7(July 6, 2012)
- ISSN：1535-3907
文摘
Aberrations in skin morphology and functionality can cause acute and chronic skin-related diseases that are the focus of dermatological research. Mechanically induced skin suction blister fluid may serve as a potential, alternative human body fluid for quantitative mass spectrometry (MS)-based proteomics in order to assist in the understanding of the mechanisms and causes underlying skin-related diseases. The combination of abundant-protein removal with iTRAQ technology and multidimensional fractionation techniques improved the number of identified protein groups. A relative comparison of a cohort of 8 healthy volunteers was thus recruited in order to assess the net variability encountered in a healthy scenario. The technology enabled the identification, to date, of the highest number of reported protein groups (739) with concomitant relative quantitative data for over 90% of all proteins with high reproducibility and accuracy. The use of iTRAQ 8-plex resulted in a 66% decrease in protein identifications but, despite this, provided valuable insight into interindividual differences of the healthy control samples. The geometric mean ratio was close to 1 with 95% of all ratios ranging between 0.45 and 2.05 and a calculated mean coefficient of variation of 15.8%, indicating a lower biological variance than that reported for plasma or urine. By applying a multistep sample processing, the obtained sensitivity and accuracy of quantitative MS analysis demonstrates the prospective value of the approach in future research into skin diseases.