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Establishment and Clinical Application of a Highly Sensitive Time-Resolved Fluorescence Immunoassay for Tumor-Associated Trypsinogen-2

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Abstract

To establish a rapid and highly sensitive assay for tumor-associated trypsinogen-2 (TAT-2) based on the time-resolved fluorescence immunoassay (TRFIA) and evaluate its potential clinical value in patients with lung cancer. The double-antibody sandwich method was used in detecting TAT-2 antigen concentrations, and two types of TAT-2 antibodies (coating antibodies and Eu3+ labeled antibodies) were used. A TAT-2–TRFIA method was then established, evaluated, and used in detecting the serum TAT-2 levels of healthy subjects and patients with lung cancer. The linear range of the TAT-2–TRFIA method was 1.53–300 ng/mL, the intra-assay coefficient of variation (CV) were between 1.67% and 8.42%, and the inter-assay CV were between 4.29% and 11.44%. The recovery rates of TAT-2–TRFIA were between 99.17% and 107.06%. The cross-reactivities of trypsin and T-cell immunoglobulin mucin 3 were 0.02% and 0.82%, respectively. The serum TAT-2 levels of patients with lung cancer were higher than those of healthy subjects (P < 0.001). Combined with TAT-2, the sensitivity and specificity of CEA and CA-125 for lung cancer improved significantly.

Conclusion: We successfully established a highly sensitive TAT-2–TRFIA method, which was able to facilitate the timely diagnosis of lung cancer.

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Availability of Data and Material

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

References

  1. Pham D, Bhandari S, Pinkston C, Oechsli M, Kloecker G (2020) Lung Cancer Screening Registry Reveals Low-dose CT Screening Remains Heavily Underutilized. Clin Lung Cancer 21(3):e206–e211

    Article  Google Scholar 

  2. Ruchalski K, Gutierrez A, Genshaft S, Abtin F, Suh R (2016) The evidence for low-dose CT screening of lung cancer. Clin Imaging 40(2):288–295

    Article  Google Scholar 

  3. Shen H (2018) Low-dose CT for lung cancer screening: opportunities and challenges. Front Med 12(1):116–121

    Article  Google Scholar 

  4. Collins LG, Haines C, Perkel R, Enck RE (2007) Lung cancer: diagnosis and management. Am Fam Physician 75(1):56–63

    PubMed  Google Scholar 

  5. Maclay JD, Farley JM, McCowan C, Tweed C, Milroy R (2017) Obtaining tissue diagnosis in lung cancer patients with poor performance status and its influence on treatment and survival. Respir Med 124:30–35

    Article  Google Scholar 

  6. Nanavaty P, Alvarez MS, Alberts WM (2014) Lung cancer screening: advantages, controversies, and applications. Cancer Control 21(1):9–14

    Article  Google Scholar 

  7. Jiang JC, Zhang Y (2020) Serological antibody testing in the COVID-19 pandemic: their molecular basis and applications. Biochem Soc Trans 48(6):2851–2863

    Article  CAS  Google Scholar 

  8. Lau A, Chen S, Sleiman S, Sorrell T (2009) Current status and future perspectives on molecular and serological methods in diagnostic mycology. Future Microbiol 4(9):1185–1222

    Article  CAS  Google Scholar 

  9. Li X, Asmitananda T, Gao L, Gai D, Song Z, Zhang Y, Ren H, Yang T, Chen T, Chen M (2012) Biomarkers in the lung cancer diagnosis: a clinical perspective. Neoplasma 59(5):500–507

    Article  CAS  Google Scholar 

  10. Wang R, Wang G, Zhang N, Li X, Liu Y (2013) Clinical evaluation and cost-effectiveness analysis of serum tumor markers in lung cancer. Biomed Res Int 2013:195692

    PubMed  PubMed Central  Google Scholar 

  11. Burt RW, Ratcliffe JG, Stack BH, Cuthbert J, Kennedy RS, Corker CS, Franchimont P, Spilg WG, Stimson WH (1978) Serum biochemical markers in lung cancer. Br J Cancer 37(5):714–717

    Article  CAS  Google Scholar 

  12. Isaksson S, Jönsson P, Monsef N, Brunnström H, Bendahl PO, Jönsson M, Staaf J, Planck M (2017) CA 19–9 and CA 125 as potential predictors of disease recurrence in resectable lung adenocarcinoma. PLoS ONE 12(10):e0186284

    Article  Google Scholar 

  13. Grunnet M, Sorensen JB (2012) Carcinoembryonic antigen (CEA) as tumor marker in lung cancer. Lung Cancer 76(2):138–143

    Article  CAS  Google Scholar 

  14. Itkonen O (2010) Human trypsinogens in the pancreas and in cancer. Scand J Clin Lab Invest 70(2):136–143

    Article  CAS  Google Scholar 

  15. Kawano N, Osawa H, Ito T, Nagashima Y, Hirahara F, Inayama Y, Nakatani Y, Kimura S, Kitajima H, Koshikawa N et al (1997) Expression of gelatinase A, tissue inhibitor of metalloproteinases-2, matrilysin, and trypsin(ogen) in lung neoplasms: an immunohistochemical study. Hum Pathol 28(5):613–622

    Article  CAS  Google Scholar 

  16. Kohn EC, Liotta LA (1995) Molecular insights into cancer invasion: strategies for prevention and intervention. Cancer Res 55(9):1856–1862

    CAS  PubMed  Google Scholar 

  17. Koivunen E, Ristimäki A, Itkonen O, Osman S, Vuento M, Stenman UH (1991) Tumor-associated trypsin participates in cancer cell-mediated degradation of extracellular matrix. Cancer Res 51(8):2107–2112

    CAS  PubMed  Google Scholar 

  18. Sorsa T, Salo T, Koivunen E, Tyynelä J, Konttinen YT, Bergmann U, Tuuttila A, Niemi E, Teronen O, Heikkilä P et al (1997) Activation of type IV procollagenases by human tumor-associated trypsin-2. J Biol Chem 272(34):21067–21074

    Article  CAS  Google Scholar 

  19. Yang X, Ye Y, Wang T, Li M, Yu L, Xia M, Qian J, Hu Z (2019) Eu(3+) /Sm(3+) dual-label time-resolved fluoroimmunoassay for measurement of hepatitis C virus antibodies. J Clin Lab Anal 33(2):e22659

    Article  Google Scholar 

  20. Zhang Q, Huang B, Liu X, Liu B, Zhang Y, Zhang Z, Hua J, Fan Y, Hu L, Meng M et al (2017) Ultrasensitive quantitation of anti-phospholipase A2 receptor antibody as a diagnostic and prognostic indicator of idiopathic membranous nephropathy. Sci Rep 7(1):12049

    Article  Google Scholar 

  21. Huang B, Xiao H, Zhang X, Zhu L, Liu H, Jin J (2006) Ultrasensitive detection of pepsinogen I and pepsinogen II by a time-resolved fluoroimmunoassay and its preliminary clinical applications. Anal Chim Acta 571(1):74–78

    Article  CAS  Google Scholar 

  22. Koivunen E, Huhtala ML, Stenman UH (1989) Human ovarian tumor-associated trypsin. Its purification and characterization from mucinous cyst fluid and identification as an activator of pro-urokinase. J Biol Chem 264(24):14095–14099

    Article  CAS  Google Scholar 

  23. Koivunen E, Saksela O, Itkonen O, Osman S, Huhtala ML, Stenman UH (1991) Human colon carcinoma, fibrosarcoma and leukemia cell lines produce tumor-associated trypsinogen. Int J Cancer 47(4):592–596

    Article  CAS  Google Scholar 

  24. Lukkonen A, Sorsa T, Salo T, Tervahartiala T, Koivunen E, Golub L, Simon S, Stenman UH (2000) Down-regulation of trypsinogen-2 expression by chemically modified tetracyclines: association with reduced cancer cell migration. Int J Cancer 86(4):577–581

    Article  CAS  Google Scholar 

  25. Stenman M, Paju A, Hanemaaijer R, Tervahartiala T, Leminen A, Stenman UH, Konttinen YT, Sorsa T (2003) Collagenases (MMP-1, -8 and -13) and trypsinogen-2 in fluid from benign and malignant ovarian cysts. Tumour Biol 24(1):9–12

    Article  CAS  Google Scholar 

  26. Kitamura H, Oosawa Y, Kawano N, Kameda Y, Hayashi H, Nakatani Y, Udaka N, Ito T, Miyazaki K (1999) Basement membrane patterns, gelatinase A and tissue inhibitor of metalloproteinase-2 expressions, and stromal fibrosis during the development of peripheral lung adenocarcinoma. Hum Pathol 30(3):331–338

    Article  CAS  Google Scholar 

  27. Kimland M, Russick C, Marks WH, Borgström A (1989) Immunoreactive anionic and cationic trypsin in human serum. Clin Chim Acta 184(1):31–46

    Article  CAS  Google Scholar 

  28. Huang B, Yang X, Zhang W, Wu J, Liu P, Hu Z, Wang T (2020) Receptor antibody time-resolved A2 phospholipase bead immunochromatography and its application in idiopathic membranous nephropathy. J Clin Lab Anal 34(12):e23508

  29. Chen M, Wang L, Wang Y, Zhou X, Liu X, Chen H, Huang B, Hu Z (2020) Soluble Tim3 detection by time-resolved fluorescence immunoassay and its application in membranous nephropathy. J Clin Lab Anal 34(6):e23248

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

Funding was provided by the Social Development Fund of Zhejiang Province (No. LGF20H200008), the Key Research and Development Program of Zhejiang Province (No. 2020C03066), “Two Hundreds” Young and Middle-aged Medical Key Talent Project of Wuxi (No. BJ2020027), the Opening Project of Zhejiang Provincial Preponderant and Characteristic Subject of Key University (Traditional Chinese Pharmacology), Key Research and Development Project of Hangzhou (No.202004A23).

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Author notes

  1. Xindong Chen, Jianfeng Hong and Han Zhao contributed equally to the work.

Authors and Affiliations

  1. College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, China

    Xindong Chen, Zhongyi Xiang, Yuan Qin, Xiumei Zhou, Yigang Wang, Pengguo Xia & Biao Huang

  2. Affiliated Xiaoshan Hospital, Hangzhou Normal University, Hangzhou, China

    Jianfeng Hong, Liping Zheng & Hongming Fang

  3. Coll Pharm, Lab Med Plant Biotechnol, Zhejiang Chinese Med Univ, Hangzhou, Zhejiang, 311402, China

    Pengguo Xia

  4. The Affiliated Wuxi No.2 People’s Hospital of Nanjing Medical University, Nanjing, China

    Han Zhao & Yingwei Zhu

Authors
  1. Xindong Chen
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  2. Jianfeng Hong
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  4. Zhongyi Xiang
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Contributions

Xindong Chen: Conceptualization, Methodology, Writing-Original Draft. Jianfeng Hong, Han Zhao: Software, Formal analysis. Zhongyi Xiang: Data Curation, Visualization. Yuan Qin, Xiumei Zhou, Yigang Wang, Liping Zheng, Biao Huang: Reviewing. Biao Huang, Pengguo Xia provided the funds. Biao Huang, Yingwei Zhu, Hongming Fang: Project administration and Supervision. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Hongming Fang, Yingwei Zhu or Biao Huang.

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Ethical Approval

The overall design of this research was approved by Zhejiang Xiaoshan Hospital, and this research was approved by the ethics committee of Zhejiang Xiaoshan Hospital. Ethic number is 2020–011.

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Informed consent was obtained from all individual participants included in the study.

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Patients signed informed consent regarding publishing their data and photographs.

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Chen, X., Hong, J., Zhao, H. et al. Establishment and Clinical Application of a Highly Sensitive Time-Resolved Fluorescence Immunoassay for Tumor-Associated Trypsinogen-2. J Fluoresc 32, 1501–1507 (2022). https://doi.org/10.1007/s10895-022-02950-1

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  • DOI: https://doi.org/10.1007/s10895-022-02950-1

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