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Article

Comprehensive Proteomic Analysis of the Human Amniotic Fluid Proteome: Gestational Age-Dependent Changes

Supporting Info

John-Edward A. Michaels, Surendra Dasari,^‡ Leonardo Pereira,^† Ashok P. Reddy, Jodi A. Lapidus, Xinfang Lu,^‡ Thomas Jacob, Archana Thomas, Matthew Rodland,^‡ Charles T. Roberts, Jr.,^‡ Michael G. Gravett,^§ and Srinivasa R. Nagalla*^§^‡

Departments of Pediatrics and Obstetrics and Gynecology, Oregon Health and Science University, Portland, Oregon 97239, Department of Obstetrics and Gynecology, University of Washington, Seattle, Washington 98101, and ProteoGenix, Inc., Portland, Oregon 97213

J. Proteome Res., 2007, 6 (4), pp 1277–1285

DOI: 10.1021/pr060543t

Publication Date (Web): March 21, 2007

ProteoGenix, Inc.

‡

Department of Pediatrics, Oregon Health and Science University.

†

Department of Obstetrics and Gynecology, Oregon Health and Science University.

University of Washington.

To whom correspondence should be addressed. E-mail, nagallas@ ohsu.edu; tel, 503-494-1928; fax, 503-494-4821.

Abstract

Amniotic fluid (AF) is a significant contributor to fetal health and constitutes a potential rich source of biomarkers for diagnosis of maternal and fetal disorders. In this study, we performed a comprehensive survey of the proteins expressed in AF, combining gel and liquid-based fractionation approaches coupled with LC−MS/MS analysis. Two-dimensional Liquid Chromatography (2D-LC) analysis identified 118 nonredundant proteins with high confidence. One- and two-dimensional gel electrophoresis and in-gel digestion identified 101 proteins. Combining both sets resulted in 219 proteins, of which 96 are unique to AF; 70, 18, and 35 proteins are present in serum, cervico-vaginal fluid, and all three fluids, respectively. Fluorescence two-dimensional differential in-gel electrophoresis (2D-DIGE) comparison of first-, second-, and third-trimester AF samples revealed that maximal differences in the relative abundance of AF proteins occur between the first and second trimesters. A systematic analysis of proteins present both in AF and maternal serum could lead to the development of new noninvasive diagnostic procedures to monitor fetal status.

Keywords: Amniotic fluid • Proteome • Pregnancy • Biomarkers • Aneuploidy

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