Development of a sandwich ELISA for the thrombin light chain identified by serum proteome analysis
We previously identified novel biomarker candidates in biliary tract cancer (BTC) using serum proteome analysis. Among several candidates, we focused on thrombin light chain which is a 4204 Da peptide as the most promising biomarker for BTC. To move thrombin light chain toward potential diagnostic u...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2017-08-01
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| Series: | Practical Laboratory Medicine |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352551716300403 |
| _version_ | 1811256192008192000 |
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| author | Kazuyuki Sogawa Kana Kobayashi Shintaro Kikkawa Shigetsugu Takano Hideyuki Yoshitomi Hirotaka Takizawa Masayuki Ohtsuka Hiroaki Shimizu Katsunori Furuhata Masaru Miyazaki Osamu Yokosuka Fumio Nomura |
| author_facet | Kazuyuki Sogawa Kana Kobayashi Shintaro Kikkawa Shigetsugu Takano Hideyuki Yoshitomi Hirotaka Takizawa Masayuki Ohtsuka Hiroaki Shimizu Katsunori Furuhata Masaru Miyazaki Osamu Yokosuka Fumio Nomura |
| author_sort | Kazuyuki Sogawa |
| collection | DOAJ |
| description | We previously identified novel biomarker candidates in biliary tract cancer (BTC) using serum proteome analysis. Among several candidates, we focused on thrombin light chain which is a 4204 Da peptide as the most promising biomarker for BTC. To move thrombin light chain toward potential diagnostic use, we developed an enzyme immunoassay that enables to measure serum thrombin light chain levels.Both one monoclonal antibody specific to the N-termini and one polyclonal antibody were used to develop a sandwich ELISA for thrombin light chain. The assay was evaluated by comparing the results with those obtained by the ClinProt™ system. Serum samples were obtained from 20 patients with BTC, 20 patients with BBTDs and 20 HVs using the ClinProt™ system and ELISA.The results of the established ELISA showed a positive correlation with the findings by ClinProt™ system (slope=0.3386, intercept=34.901, r2=0.9641). The performance of the ELISA was satisfactory in terms of recovery (97.9–102.5%) and within-run (1.5–4.8%) and between-day (1.9–6.7%) reproducibility. Serum thrombin light chain levels were significantly greater in BTC (176.5±47.2 ng/mL) than in BBTDs (128.6±17.4 ng/mL) and HVs (127.6±16.0 ng/mL) (p<0.001).The sandwich ELISA developed in this study will be useful for validation of the diagnostic significance of serum thrombin light chain levels in various cancers. Keywords: Thrombin light chain, Biliary tract cancer, Sandwich ELISA, Serum biomarker |
| first_indexed | 2024-04-12T17:37:18Z |
| format | Article |
| id | doaj.art-0c9449e3eeeb4323b87f3ccfd95b80ee |
| institution | Directory Open Access Journal |
| issn | 2352-5517 |
| language | English |
| last_indexed | 2024-04-12T17:37:18Z |
| publishDate | 2017-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Practical Laboratory Medicine |
| spelling | doaj.art-0c9449e3eeeb4323b87f3ccfd95b80ee2022-12-22T03:22:56ZengElsevierPractical Laboratory Medicine2352-55172017-08-0183440Development of a sandwich ELISA for the thrombin light chain identified by serum proteome analysisKazuyuki Sogawa0Kana Kobayashi1Shintaro Kikkawa2Shigetsugu Takano3Hideyuki Yoshitomi4Hirotaka Takizawa5Masayuki Ohtsuka6Hiroaki Shimizu7Katsunori Furuhata8Masaru Miyazaki9Osamu Yokosuka10Fumio Nomura11Department of Biochemistry, School of Life and Environmental Science, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara-shi, Kanagawa 252-5201, Japan; Corresponding author.Department of Biochemistry, School of Life and Environmental Science, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara-shi, Kanagawa 252-5201, JapanDepartment of Medicine and Clinical Oncology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba 260-8670, JapanDepartment of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba 260-8670, JapanDepartment of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba 260-8670, JapanKashiwado Clinic in Port-Square, Kashiwado Memorial Foundation, 1-35 Tonyachou, Chuo-ku, Chiba-shi, Chiba 260-0025, JapanDepartment of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba 260-8670, JapanDepartment of Medicine and Clinical Oncology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba 260-8670, JapanDepartment of Biochemistry, School of Life and Environmental Science, Azabu University, 1-17-71 Fuchinobe, Chuo-ku, Sagamihara-shi, Kanagawa 252-5201, JapanDepartment of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba 260-8670, JapanDepartment of Medicine and Clinical Oncology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba 260-8670, JapanDivisions of Clinical Mass Spectrometry and Clinical Genetics, Chiba University Hospital, 1-8-1 Inohana, Chuo-ku, Chiba-shi, Chiba 260-8670, JapanWe previously identified novel biomarker candidates in biliary tract cancer (BTC) using serum proteome analysis. Among several candidates, we focused on thrombin light chain which is a 4204 Da peptide as the most promising biomarker for BTC. To move thrombin light chain toward potential diagnostic use, we developed an enzyme immunoassay that enables to measure serum thrombin light chain levels.Both one monoclonal antibody specific to the N-termini and one polyclonal antibody were used to develop a sandwich ELISA for thrombin light chain. The assay was evaluated by comparing the results with those obtained by the ClinProt™ system. Serum samples were obtained from 20 patients with BTC, 20 patients with BBTDs and 20 HVs using the ClinProt™ system and ELISA.The results of the established ELISA showed a positive correlation with the findings by ClinProt™ system (slope=0.3386, intercept=34.901, r2=0.9641). The performance of the ELISA was satisfactory in terms of recovery (97.9–102.5%) and within-run (1.5–4.8%) and between-day (1.9–6.7%) reproducibility. Serum thrombin light chain levels were significantly greater in BTC (176.5±47.2 ng/mL) than in BBTDs (128.6±17.4 ng/mL) and HVs (127.6±16.0 ng/mL) (p<0.001).The sandwich ELISA developed in this study will be useful for validation of the diagnostic significance of serum thrombin light chain levels in various cancers. Keywords: Thrombin light chain, Biliary tract cancer, Sandwich ELISA, Serum biomarkerhttp://www.sciencedirect.com/science/article/pii/S2352551716300403 |
| spellingShingle | Kazuyuki Sogawa Kana Kobayashi Shintaro Kikkawa Shigetsugu Takano Hideyuki Yoshitomi Hirotaka Takizawa Masayuki Ohtsuka Hiroaki Shimizu Katsunori Furuhata Masaru Miyazaki Osamu Yokosuka Fumio Nomura Development of a sandwich ELISA for the thrombin light chain identified by serum proteome analysis Practical Laboratory Medicine |
| title | Development of a sandwich ELISA for the thrombin light chain identified by serum proteome analysis |
| title_full | Development of a sandwich ELISA for the thrombin light chain identified by serum proteome analysis |
| title_fullStr | Development of a sandwich ELISA for the thrombin light chain identified by serum proteome analysis |
| title_full_unstemmed | Development of a sandwich ELISA for the thrombin light chain identified by serum proteome analysis |
| title_short | Development of a sandwich ELISA for the thrombin light chain identified by serum proteome analysis |
| title_sort | development of a sandwich elisa for the thrombin light chain identified by serum proteome analysis |
| url | http://www.sciencedirect.com/science/article/pii/S2352551716300403 |
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