Development of immunocomplex pretreatment method for mass spectrometry (MS, Immuno-MS) which is applicable to clinical testing

• Project/Area Number: 15K08611
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Chiba University
• Principal Investigator: Nishimura Motoi 千葉大学, 医学部附属病院, 講師 (80400969)
• Co-Investigator(Kenkyū-buntansha): 佐藤 守 千葉大学, 医学部附属病院, 特任准教授 (20401002)
• Co-Investigator(Renkei-kenkyūsha): KODERA Yoshio 北里大学, 理学部, 准教授 (60265733)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2015: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
• Keywords: 質量分析 / ビタミンD / 細菌同定 / 病態検査学 / 臨床検査 / ペプチドミクス / プロテオミクス / 臨床検査医学 / イムノアッセイ

Outline of Final Research Achievements

Pretreatment methods for mass spectrometry to separate and measure antigen (peptide / protein) from immunocomplex without LC (liquid chromatography) was studied. Specifically, denaturation treatment and ultrafiltration were carried out to separate antigens. Even if MALDI-MS (mass spectrometry by matrix assisted laser desorption / ionization method) ,which is relatively resistant to contaminants in samples is used, it is difficult to obtain sufficient reproducibility for detection data when deviating from the ideal condition.
We further conducted mass spectrometry research on tumor secretion, liver disease marker, vitamin D metabolite etc. and published peer reviewed studies. In MALDI - MS in the bacterial identification test of blood culture specimens, good results of clinical examination level were obtained by pretreatment method not dependent on LC by using special polymer.

Research Products

• [Journal Article] Assessing the accuracy of variant detection in cost-effective gene panel testing by next-generation sequencing (2018)
  • Author(s): Fujiki R, Yoshida A, Ohara O, Morisaki H, Nishimura M, Ichikawa T, Morisaki T, YAO Y, Tanaka T, Matsushita K, Ikeda M, Maeda A.
  • Journal Title: The Journal of Molecular Diagnostics Volume: in press
  • Peer Reviewed
• [Journal Article] Assessment by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry of the Effects of Preanalytical Variables on Serum Peptidome Profiles Following Long-Term Sample Storage. (2018)
  • Author(s): Tsuchida S, Satoh M, Umemura H, Sogawa K, Takiwaki M, Ishige T, Miyabayashi Y, Iwasawa Y, Kobayashi S, Beppu M, Nishimura M, Kodera Y, Matsushita K, Nomura F
  • Journal Title: Proteomics. Clinical applications Volume: 15 Issue: 3 Pages: 47-53
  • DOI: 10.1002/prca.201700047
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Variant in C-terminal region of intestinal alkaline phosphatase associated with benign familial hyperphosphatasaemia. (2018)
  • Author(s): Ishige T, Itoga S, Utsuno E, Nishimura M, Yoshikawa M, Kato N, Matsushita K, Yokosuka O, Nomura F
  • Journal Title: Journal of medical genetics Volume: 10 Issue: 10 Pages: 47-53
  • DOI: 10.1136/jmedgenet-2017-104964
  • Peer Reviewed / Open Access
• [Journal Article] Surveillance evaluation of the standardization of assay values for serum total 25-hydroxyvitamin D concentration in Japan. (2018)
  • Author(s): Ihara H, Kiuchi S, Ishige T, Nishimura M, Matsushita K, Satoh M, Nomura F, Yamashita M, Kitajima I, Tsugawa N, Okano T, Hirota K, Miura M, Totani M, Hashizume N; Nutrition Division, Japan Society of Clinical Chemistry.
  • Journal Title: Ann Clin Biochem. Volume: Epub ahead of print Issue: 6 Pages: 0-0
  • DOI: 10.1177/0004563218765570
  • Peer Reviewed / Open Access
• [Journal Article] Improved sensitivity of serum/plasma 1α,25-dihydroxyvitamin D quantification by DAPTAD derivatization (2017)
  • Author(s): Ishige Takayuki, Satoh Mamoru, Ogawa Shoujiro, Nishimura Motoi, Matsushita Kazuyuki, Higashi Tatsuya, Nomura Fumio
  • Journal Title: Clinica Chimica Acta Volume: 473 Pages: 173-179
  • DOI: 10.1016/j.cca.2017.08.033
  • Peer Reviewed / Open Access
• [Journal Article] Applications of copolymer for rapid identification of bacteria in blood culture broths using matrix-assisted laser desorption ionization time-of-flight mass spectrometry. (2017)
  • Author(s): Ashizawa K, Murata S, Terada T, Ito D, Bunya M, Watanabe K, Teruuchi Y, Tsuchida S, Satoh M, Nishimura M, Matsushita K, Sugama Y, Nomura F
  • Journal Title: Journal of microbiological methods Volume: 139 Pages: 54-60
  • DOI: 10.1016/j.mimet.2017.04.013
  • Peer Reviewed / Open Access
• [Journal Article] A novel KCNQ1 nonsense variant in the isoform-specific first exon causes both jervell and Lange-Nielsen syndrome 1 and long QT syndrome 1: a case report. (2017)
  • Author(s): Nishimura M, Ueda M, Ebata R, Utsuno E, Ishii T, Matsushita K, Ohara O, Shimojo N, Kobayashi Y, Nomura F
  • Journal Title: BMC medical genetics Volume: 18 Issue: 1 Pages: 66-71
  • DOI: 10.1186/s12881-017-0430-7
  • Peer Reviewed / Open Access
• [Journal Article] Rapid and cost-effective high-throughput sequencing for identification of germline mutations of BRCA1 and BRCA2 (2017)
  • Author(s): Ahmadloo S, Nakaoka H, Hayano T, Hosomichi K, You H, Utsuno E, Sangai T, Nishimura M, Matsushita K, Hata A, Nomura F, Inoue I
  • Journal Title: J Hum Genetics Volume: 印刷中 Issue: 5 Pages: 561-567
  • DOI: 10.1038/jhg.2017.5
  • NAID: 40021210379
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Serum cytokine and chemokine profiles in patients with juvenile muscular atrophy of distal upper extremity (Hirayama disease). (2017)
  • Author(s): Beppu M, Sawai S, Misawa S, Mori M, Ito S, Sogawa K, Nishimura M, Matsushita K, Nomura F, Kuwabara S.
  • Journal Title: J Neuroimmunol Volume: 302 Pages: 20-22
  • DOI: 10.1016/j.jneuroim.2016.11.011
  • Peer Reviewed / Open Access
• [Journal Article] Development and validation of the simultaneous measurement of four vitamin D metabolites in serum by LC-MS/MS for clinical laboratory applications (2016)
  • Author(s): Mamoru Satoh, Takayuki Ishige, Shoujiro Ogawa, Motoi Nishimura, Kazuyuki Matsushita, Tatsuya Higashi, Fumio Nomura
  • Journal Title: Anal Bioanal Chem Volume: 408 Issue: 27 Pages: 7617-7627
  • DOI: 10.1007/s00216-016-9821-4
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Combined Secretomics and Transcriptomics Revealed Cancer-Derived GDF15 is Involved in Diffuse-Type Gastric Cancer Progression and Fibroblast Activation. (2016)
  • Author(s): Ishige T, Nishimura M, Satoh M, Fujimoto M, Fukuyo M, Semba T, Kado S, Tsuchida S, Sawai S, Matsushita K, Togawa A, Matsubara H, Kaneda A, Nomura F
  • Journal Title: Sci Rep. Volume: 6 Issue: 1 Pages: 21681-21681
  • DOI: 10.1038/srep21681
  • Peer Reviewed / Open Access
• [Journal Article] Coupling CDH17 and CLDN18 markers for comprehensive membrane-targeted detection of human gastric cancer. (2016)
  • Author(s): Matsusaka K, Ushiku T, Urabe M, Fukuyo M, Abe H, Ishikawa S, Seto Y, Aburatani H, Hamakubo T, Kaneda A, Fukayama M.
  • Journal Title: Oncotarget Volume: 7 Issue: 51 Pages: 64168-64181
  • DOI: 10.18632/oncotarget.11510
  • Peer Reviewed / Open Access
• [Journal Article] Fibrinogen alpha C chain 5.9 kDa fragment (FIC5.9), a biomarker for various pathological conditions, is produced in post-blood collection by fibrinolysis and coagulation factors (2016)
  • Author(s): Kikuchi W, Nishimura M, Kuga T, Tsuchida S, Saito T, Satoh M, Noda K, Kodera Y, Tomonaga T, Nomura F
  • Journal Title: Clinical Proteomics Volume: 13 Issue: 1 Pages: 27-27
  • DOI: 10.1186/s12014-016-9129-6
  • Peer Reviewed / Open Access
• [Journal Article] Familial lamin A/C mutation cardiomyopathy with arrhythmia substrate detected by cardiac magnetic resonance imaging and electroanatomical mapping. (2016)
  • Author(s): Nishi T, Takaoka H, Funabashi N, Nishimura M, Ohara O, Makiyama T, Ueda M, Kajiyama T, Kobayashi Y.
  • Journal Title: International Journal of Cardiology Volume: 209 Pages: 248-252
  • DOI: 10.1016/j.ijcard.2016.02.030
  • Peer Reviewed / Open Access
• [Journal Article] Detection of Ubiquitinated Dermcidin in Gingival Crevicular Fluid in Periodontal Disease (2015)
  • Author(s): Sachio Tsuchida , Mamoru Satoh, Masaki Takiwaki, Michiru Wakabayashi, Takayuki Ishige, Minako Beppu, Motoi Nishimura, Kazuyuki Matsushita, Fumio Nomura
  • Journal Title: International Journal of Peptide Research and Therapeutics Volume: 1 Pages: 1-5
  • Peer Reviewed / Open Access
• [Journal Article] Identification of Nocardia species using matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry (2015)
  • Author(s): Segawa S, Nishimura M, Sogawa K, Tsuchida S, Murata S, Watanabe M, Matsushita K, Kamei K and Nomura F.
  • Journal Title: Clinical Proteomics Volume: 12 Issue: 1 Pages: 1-1
  • DOI: 10.1186/s12014-015-9078-5
  • Peer Reviewed / Open Access
• [Journal Article] Assessment of Vitamin D Metabolism by LC-MS/MS (2015)
  • Author(s): Ishige T, Satoh M, Ogawa S, Nishimura M, Higashi T, Nomura F.
  • Journal Title: Rinsho Byori. Volume: 63 Pages: 457-464
  • NAID: 40020474101
• [Journal Article] Laboratory medicine and sex differences (2015)
  • Author(s): Nishimura M, Nomura F.
  • Journal Title: Nihon Rinsho Volume: 73 Pages: 587-600
  • NAID: 40020398301
• [Presentation] 血液培養陽性検体よりの複数菌由来の核酸検出 (2018)
  • Author(s): 西村 基
  • Organizer: 第29回日本臨床微生物学会総会・学術集会
• [Presentation] LQT5(KCNE1)D85Nの合併による重症化が推定されたLQT2の1家系 (2017)
  • Author(s): 西村基,齋藤直樹,江畑亮太,奥主健太郎,上田希彦,宇津野恵美,別府美奈子,松下一之,小原収,田中知明,小林欣夫,下条直樹,市川智彦
  • Organizer: 第24回日本遺伝子診療学会大会
• [Presentation] 新規慢性肝疾患マーカーFIC5.9( Fibrinogen alpha C chain5.9kD fragment ) は凝固線溶の変動を反映する (2017)
  • Author(s): 西村基,菊地渉,佐藤守,松下一之,小寺義男,田中知明, 野村文夫
  • Organizer: 第57回日本臨床化学会年次学術集会
• [Presentation] 慢性肝疾患で低下する新規マーカーFIC5.9(Fibrinogen αC chain 5.9 kD fragment) の健常時産生メカニズム (2017)
  • Author(s): 西村基
  • Organizer: 第64回日本臨床検査医学会学術集会・第29回世界病理臨床検査医学会連合
  • Int'l Joint Research
• [Presentation] qPCR-HRM法およびNGSにより同定したAPC遺伝子Exon8欠損による家族性大腸腺腫症家系 (2016)
  • Author(s): 松下一之、糸賀栄、石毛崇之、西村基、別府美奈子、内垣洋祐、宇津野恵美、澤部祐司、野村文夫
  • Organizer: 日本臨床検査自動化学第48回大会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-09-23
• [Presentation] Long-PCRと大量並列シーケンシングによる家族性地中海熱の原因遺伝子MEFVの遺伝子学的検査 (2016)
  • Author(s): 石毛崇之、糸賀栄、北村浩一、西村基、松下一之、野村文夫
  • Organizer: 日本臨床検査自動化学第48回大会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-09-22
• [Presentation] 質量分析技術による「ゲノム・フリー」なApolipoprotein Eタイピング（迅速ApoE セロタイピング）を目指した試み (2016)
  • Author(s): 西村 基
  • Organizer: 第63回日本臨床検査医学会学術集会
  • Place of Presentation: 神戸国際会議場（兵庫県神戸市）
  • Year and Date: 2016-09-02
• [Presentation] ゲノム解析における偶発的所見に対する遺伝カウンセリングの必要性と課題 (2016)
  • Author(s): 松下一之、西村基、別府美奈子、糸賀栄、石毛崇之、北村浩一、宇津野恵美、澤部祐司、野村文夫
  • Organizer: 第40回日本遺伝カウンセリング学会学術集会/ The 13th International Congress of Human Genetics
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2016-04-06
  • Int'l Joint Research
• [Presentation] 遺伝性脊髄小脳変性症の複数の遺伝子変異を合併した症例における遺伝カウンセリング (2016)
  • Author(s): 別府美奈子、澤井摂、宇津野恵美、石毛崇之、糸賀栄、磯瀬沙希里、新井公人、鈴木陽一、吉川由利子、西村基、野村文夫、桑原聡、松下一之
  • Organizer: 第40回日本遺伝カウンセリング学会学術集会/ The 13th International Congress of Human Genetics
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2016-04-06
  • Int'l Joint Research
• [Presentation] ミトコンドリアゲノム全周解析のための簡便な前処理法 (2016)
  • Author(s): 西村 基、西村里美、姚躍、石毛崇之、糸賀栄、松下一之、野村文夫
  • Organizer: 第40回日本遺伝カウンセリング学会学術集会/ The 13th International Congress of Human Genetics
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2016-04-05
  • Int'l Joint Research
• [Presentation] 大学検査部に適した「質量分析手法を用いた次世代臨床検査」とは (2015)
  • Author(s): 西村基
  • Organizer: 日本臨床検査医学会
  • Place of Presentation: 長良川国際会議場（岐阜県岐阜市）
  • Year and Date: 2015-11-19
  • Invited
• [Presentation] 自動分析法による血清25ヒドロキシビタミンD測定値の標準化：SRM 972aを校正に用いた測定 (2015)
  • Author(s): 西村 基、佐藤 守、石毛崇之 、東 達也、 野村 文夫
  • Organizer: 日本臨床化学会
  • Place of Presentation: 大阪大学コンベンションセンター（大阪府吹田市）
  • Year and Date: 2015-10-30
  • Invited
• [Presentation] QT延長症候群遺伝子検査の次世代シーケンサーを用いた立ち上げとこれにより変異同定に至った Jervell and Lange-Nielsen Syndrome(JLNS) の一家系 (2015)
  • Author(s): 西村基 宇津野 恵美 上田 希彦 江畑 亮太 内海 紀子 糸賀 栄 小原 令子 別府 美奈子 松下 一之 小原 収 野村 文夫
  • Organizer: 日本人類遺伝学会
  • Place of Presentation: タワーホール船堀（東京都江戸川区）
  • Year and Date: 2015-10-14
• [Presentation] 保険収載された遺伝子検査・遺伝カウンセリング加算と次世代シーケンサー (2015)
  • Author(s): 西村基
  • Organizer: 日本遺伝カウンセリング学会
  • Place of Presentation: 三井ガーデンホテル千葉(千葉県千葉市)
  • Year and Date: 2015-06-26