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ポスター発表

第2日　5月16日（水）　　ポスター会場

2P-04　　PDF

SWATH法によるウシ血清タンパク質の網羅的な定量法の開発―再現性と定量性の評価―

(¹近大生物理工・ ²近大院生物工・ ³近大先技総研)
牧野加奈子¹・ ^o西端智也²・ 山脇瑳也花¹・ 池上春香¹・ 松橋珠子³・ 永井宏平^1,2・ 松本和也^1,2

Intramuscular fat (IMF) content strongly affects the economic value of beef cattle in Japan. In order to develop serum biomarkers to diagnosis the IMF content, we are planning a large scale proteomic analysis of beef cattle serum using SWATH. In this study, we performed SWATH of bovine serum in a fixed window (FIX) and a variable window (VAR) methods, and compared their reproducibility and quantitative performance. Quantitative linearity was also evaluated using spiked-in peptides. Proteins in the bovine serum were trypsinized and the resulting peptides were analyzed. Up to 172 proteins were successfully quantified by SWATH. In the five-times SWATH measurements, 79.7%, and 66.6% of the proteins were quantified with CV < 10% in FIX and VAR, respectively, indicating the higher reproducibility of FIX. In the three-times SWATH measurements in the same day and with the same ion library, number of quantified proteins were not different between the methods, but number of quantified proteins with CV < 10% or 20% were larger in FIX. These results suggested that the FIX method is suitable for quantitative proteomics of bovine serum. Quantification of spiked-in peptides by FIX showed high quantitative linearity in the peptide amount range of 10 fmol to 10 pmol.