iTRAQTM試薬とQqTOFテクノロジーを用いた転移性乳癌の早期バイオマーカー探索

Masato AOSHIMA
2006 Journal of the Mass Spectrometry Society of Japan  
QQ ̳̲̱ ̱ῸΎ̱ῢῧῌ ´QQQ῭̲´̱̱̳ῲ̳̲ῥ̰ῠQQQ῭ῧ῞̰ῌ ̮ῚῦQQQῥ̰̯ῡQ̰ῦΐQ ῩQ̰QῦQQQQ῭̱̯ΰ῝̯̲̱ ῑ̱ῑQQῘ῍΅ῥQῬΎῡ̮Ὺ῍ ´QQQ῭ῴῑ̳ΰ῝̯̲̱ ῑ̱ῑ QQῧῌ ̲̱ ῑ̱ῑΰῤῪΏ̳῾̱QῦQ'῭ῖQ῟Ὺ "discovery"ῌ ̯ῦΏ̳῾̱QῘQῷQῢῤ̮Ίΰ῭Qῥ῟Ὺ "verification"ῌ ̮Q̮ῢῦQQQQ῭Q̱῟Ὺ "validation"ῌ ´QῢῦQῗ῭̱̯ΰ῝̯ "clinical" ῦῸ̲ῑῷῥΎ̲῟ ῪΊΰῘῢῙῪ῍ QQῦῩ̰ῢῧῌ QῳΏῦQῌ QῙῦQΰῢῘ̰ΎῪῤQῦ̲̱ ῑ̱ῑ῭ῖQ ( discovery) ῟Ὺ̯̰ῥῌ ῌQQ ῲΰ̯QQ ̲̳Qῲῦ 2 Q̲ῦQῲ῭̯QQQΰ῝ῡῘ̰ΎῪ ̳̲̱ῼ̳̮Qῌ ̰̮QQῦ 2 Q̲ῦQQῢ̯Ύ ̯ΎῧῸ῝ῌ ῩQQQQῐῶῦ̳῟ῡQ̮Q̱ῢ̮Ὺ iTRAQ TM Q̱ῢ̯Ύ̯Ύ῭̳ ̳῝̯Qῌ Liquid Chromatography/Mass Spectrometry/Mass Spectrometry
more » ... ctrometry (LC/MS/MS) ῭Q̮Ίΰῢῌ Ίῦ 4 QQῢῦ̰Q῟ῼQῐ῭Q̯̯῍ ̯ ῦQQῌ ῪQQ̲ῦ̲̱ ῑ̱ῑΰῤ̰̮ῪQ'Ώ̳῾̱Q῭ῡ῟῟ῪΊΰῘῢῙῌ Ῡ̯ῌ ῡ῟Ύ̯Q'Ώ̳῾̱QῘ QῷQῢῤ̮Ίΰ῭Qῥ (verification) ῟Ὺ̯̰ῌ MIDAS TM Workflow Designer ῭ῶ̮̯ MRM (Multiple Reaction Monitoring) ῼ ̳̲̳ῦQ῎ῌ ῤ̰Ῠῥ MRM ῟῭Q̯̯ῦῢῬ̯ῡQ῟Ὺ῍ ῢ῏̯̯ 1) ̳ ̳ῦ῝῎῎ ̱̳̲̳ῑ̳Qῲΰ῝ῡ Lewis lung cancer cells (LLC-AP2) ῭ῶ̮̯῍ Ῡ̯ῌ ̯QQ ̲̳Qῲ ΰ῝ῡ ErbB2 ῭QQῩQ῟Ὺ̯QQῦ̲̳'̳̲ LLC-ErbB2-M4 ῭Q῎῝̯῍ LLC-AP2 ΰ LLC-ErbB2-M4 ῭ ̳̲̱ῼ̳̮Qῌ ̰̮QQῢ̯Ύ̯ΎῧῸ῝ῌ QQ῝̯Qῲῗ̰ΊQ῝̯Ώ̳῾̱Qῦ̮̯ 100 mg ῭̲̳ ῶ̳ῢQῒQQQῌ ̰̮ῦ ̳̲̱̳ῥQ̮̳῟ῡQ̮Q̱ῢ̮Ὺ iTRAQ TM Q̱ῥ̰Ὺ̳ ̳Q̰Q῭Q̯̯῍ 2) ̱̳ ̲̱̳ῑ῎ iTRAQ TM ̳ ̳Ύ̯̳ ̳῭Q̱ῼ̱̳QQ̱̳ ̲̱̳ῑ (2.1100 mm, 5 mm, 200 A Õ, Polysulfoethyl A column, 200 mL/min, 10῍500 mM ammonium formate, pH 3) ῥ̰̰ 40 QΎῥΎQ῝ῌ LC System ῥῧ Tempo LC ῶῸ̲ ῌ ̱̳ ῥῧ C18 ̱̳ (75 mm15 cm, LC Packings; 5῍30 acetonitrile over 30 min) ῭ῶ̮ῡ̯Ύ̯ΎῦQΎ῭ LC-MS/MS Ύῐ῝̯῍ 3) QῼΎῐῙ῎ ̱̳Qῥ NanoSprayῌ sourceῌ Ῑῧ QSTARῌ Elite ῶῸ̲ ῭ῶ̮ῡ῟῭Q̯̯῍ Ῡ̯ῌ ̲ῑΏQ̮̯ῥ̳'̳Ώ ῢ MS, MS/MS ῭Qῠ̯ῥῑ̰̮̮ῤῘ̰῟῟Ὺ Smart Information Dependent Acquisition (IDA) Qῦ ῒAnalystῌ QS Ὼ̲ῲῳ' 2.0ΐ ῭Όῶ῝̯῟ῥ̰̰̲ῑΏ῭Qῢ῝̯῍ 4) ̲ῑΏQῐ῎ Ώ̳῾̱Qῦῡ῟ΰ῟ῼQῐῥῧ Protein Pilot TM Ὼ̲ῲῳ'Ὶῦ Paragon TM Database Search ' ̳ῴ̳Ό ΰ Pro Group TM '̳ῴ̳Ό ῭ῶ̮̯῍ 5) ῩQΏ̳῾̱QῦQQ῎ QQQQῢῩQῥῴQῤQῘQ̰Ύ̯ῪQQῦΏ̳῾̱Qῥῖ̮ῡῌ ̯ῦῩQ῭῭Qῢ ῏̯ῺQ (Masato ADH=>B6); ' ̳̲̲̱ῶῸ̲ Όῷ ῾̳QQQQ ῒ̮104῍0032 ̯Q̯ῚQQῨ̱ 4῍5῍4 da Vinci QQ 1 Qΐ῏
doi:10.5702/massspec.54.214 fatcat:pezjex3cf5dyfowyqup6yksskq