Efficient prefractionation of lowabundance proteins in human plasma and construction of a twodimensional map 201

SangYun Cho, Eun-Young Lee,Joon SeokLee, Hye-YoungKim,Jae Myun Park, Min-SeokKwon, Young-Kew Park, Hyoung-Joo Lee„ Min-JungKang,Jin Young Kim, Jong Shin Yoo, SungJin Park, Jin Won Cho, Hyon-Suk Kim and Young-Ki Paik

9.1 Introduction 202

9.2 Materials and methods 203

9.2.1 Plasma sample preparation 203

9.2.2 Depletion ofmajor abundance proteins with an immunoaffinity column 203

9.2.4 Identification of proteins by MS 204

9.2.5 Fractionation of the plasma samples by FFE 204

9.2.7 Bioinformatics 206

9.3 Results and discussion 206

9.3.1 2-DE map of human plasma devoid of high-abundance proteins 206

9.3.2 Expression of different anticoagulant-treated plasma 224

9.3.3 FFE/1-DE/nanoLC-MS/MS and 2-DE/MALDI-TOF 225

9.4 Concluding remarks 229

9.5 References 229

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