Skip to main content
SpringerLink
Log in

Study on a new high affinity anti-glypicans-3 antibody in diagnosis of early hepatocellular carcinoma by differential pulse voltammetry

  • Original Paper
  • Published:
Journal of Solid State Electrochemistry Aims and scope Submit manuscript

Abstract

A novel electrochemical immunoassay for glypicans-3 (GPC3) was established based on its new high affinity antibody (Ig). In this study, we used glass carbon film (GCF) as carrier for immobilizing Ig and avoiding the high electron transfer resistance (Ret) in electrochemical immunoassay sensor (Ig-Biotin-GR/GCE) because many materials such as bovine serum albumin (BSA), avidin, biotin, Ig, GPC3, and ALP were covered on. The Ig was combined with GCP3 labeled alkaline phosphatase (ALP-GPC3) which catalyzes p-nitro phenyl phosphate (PNPP) into p-nitro phenol (PNP). In order to obtain highly sensitive electrochemical signals of the PNP and further determine ALP-GPC3, the GR/GCE modified polylysine (PLYS/GR/GCE) was fabricated and used for detecting the PNP. According to the principle of competitive inhibition, less ALP-GPC3 would be combined with the Ig in the presence of GPC3. By investigating the signals of PNP in the absence and presence of GPC3, GPC3 could be qualitatively and quantitatively analyzed. Under the optimal conditions, the peak current of PNP on the PLYS/GR/GCE was linear with the concentration of GPC3 in the range of 0.75~75 ng mL−1 and a detection limit of 0.2 ng mL−1. The recognition of GPC3 and detection of PNP were separated, which provides a new way for studying the diagnosis of liver cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Bosch FX, Ribes J, Díaz M, Cléries R (2004) Primary liver cancer: worldwide incidence and trends. Gastroenterology 127:S5–S16

    Article  Google Scholar 

  2. Thorgeirsson SS, Grisham JW (2002) Molecular pathogenesis of human hepatocellular carcinoma. Nat Genet 31:339–346

    Article  CAS  Google Scholar 

  3. Taketa K (1990) Fetoprotein: reevaluation in hepatology. Hepatology 12:1420–1432

    Article  CAS  Google Scholar 

  4. Collier J, Sherman M (1998) Elevated alphafetoprotein in benign liver diseases. Viral Hepat Rev 43:1–41

    Google Scholar 

  5. Filmus J, Church JG, Buick RN (1988) Isolation of a cDNA corresponding to a developmentally regulated transcript in rat intestine. Mol Cell Biol 8:4243–4249

    Article  CAS  Google Scholar 

  6. Filmus J, Capurro M, Rast J (2008) Glypicans. Genome Biol 9:224

    Article  Google Scholar 

  7. Hsu HC, Cheng W, Lai PL (1997) Cloning and expression of a developmentally regulated transcript MXR7 in hepatocellular carcinoma: biological significance and temporospatial distribution. Cancer Res 57:5179–5184

    CAS  Google Scholar 

  8. Zhu ZW, Friess H, Wang L, Abou-Shady M, Zimmermann A, Lander AD, Korc M, Kleeff J, Büchler MW (2001) Enhanced GPC3 expression differentiates the majority of hepatocellular carcinomas from benign hepatic disorders. Gut 48:558–564

    Article  CAS  Google Scholar 

  9. Feng J, Zhu R, Chang C, Yu L, Cao F, Zhu GH, Chen F, Xia H, Lv FD, Zhang SJ, Sun L (2016) CK19 and glypican3 expression profiling in the prognostic indication for patients with HCC after surgical resection. PLoS One 11:1–13

    Google Scholar 

  10. Capurro M, Wanless IR, Sherman M, Filmus J (2003) GPC3: a novel serum and histochemical marker for hepatocellular carcinoma. Gastroenterology 125:81–90

    Article  Google Scholar 

  11. Ho M, Kim H (2011) GPC3: a new target for cancer immunotherapy. Eur J Cancer 47:333–338

    Article  CAS  Google Scholar 

  12. Filmus J, Capurro M (2013) GPC3: a marker and a therapeutic target in hepatocellular carcinoma. FEBS J 280:2471–2476

    Article  CAS  Google Scholar 

  13. Tangkijvanich P, Chanmee T, Komtong S (2010) Diagnostic role of serum glypican-3 in differentiating hepatocellular carcinoma from non-malignant chronic liver disease and other liver cancers. J Gastroenterol Hepatol 25:129–137

    Article  CAS  Google Scholar 

  14. Capurro M, Wanless IR, Sherman M, Filmus J (2003) Glypican 3: a novel serum and histochemical marker for hepatocellular carcinoma. Gastroenterology 125:89–97

    Article  CAS  Google Scholar 

  15. Zhang QY, Chen H, Lin Z, Lin JM (2011) Chemiluminescence enzyme immunoassay based on magnetic nanoparticles for detection of hepatocellular carcinoma marker glypican-3. J Pharm Anal 1:166–174

    Article  CAS  Google Scholar 

  16. Yuan GL, He JL, Li Y, Xu WL, Gao LL, Yu C (2015) A novel ultrasensitive electrochemical immunosensor based on carboxy-endcapped conductive polypyrrole for the detection of gypican-3 in human serum. Anal Methods 7:1745–1750

    Article  CAS  Google Scholar 

  17. Huang Y, Li H, Gao T, Li G (2014) A sensitive method for protein assays using a peptide-based nano-label: human glypican-3 detection for hepatocellular carcinomas diagnosis. Analyst 139:3744–3747

    Article  CAS  Google Scholar 

  18. Jiang X, Li D, Xu X, Ying Y, Li Y, Ye Z, Wang J (2008) Immunosensors for detection of pesticide residues. Biosens Bioelectron 23:1577–1587

    Article  CAS  Google Scholar 

  19. Balkenhohl T, Lisdat F (2007) Screen-printed electrodes as impedimetric immunosensors for the detection of anti-transglutaminase antibodies in human sera. Anal Chim Acta 597:50–57

    Article  CAS  Google Scholar 

  20. Fowler JM, Stuart MC, Wong DKY (2007) Comparative study of thiolated protein G scaffolds and signal antibody conjugates in the development of electrochemical immunosensors. Biosens Bioelectron 23:633–639

    Article  CAS  Google Scholar 

  21. Tully E, Higson SP, Kennedy RO' (2008) The development of a ‘labeless’ immunosensor for the detection of Listeria monocytogenes cell surface protein, Internalin B. Biosens Bioelectron 23:906–912

    Article  CAS  Google Scholar 

  22. Lu J, Liu S, Ge S, Hu X (2012) Ultrasensitive electrochemical immunosensor based on Au nanoparticles dotted carbon nanotube-graphene composite and functionalized mesoporous materials. Biosens Bioelectron 33:29–35

    Article  Google Scholar 

  23. Jiao K, Sun W, Wang H (2001) A system of p-nitrophenylphate-alkaline phosphatase for voltammetric enzyme-linked immunoassay. Chinese J Anal Chem 29:1174–1177

    CAS  Google Scholar 

Download references

Acknowledgments

We are grateful for the financial support of the National Natural Science Foundation of China (No. 21105023), A Project of Shandong Province Higher Educational Science and Technology Program (J14LC55), the Natural Science Foundation of Shandong Province, China (ZR2014BL020, BS2013HZ027), the Shandong Province Science and Technology Development Plan Grant (2013GSF12106), and Scientific Research Foundation of Heze University (XY14KJ02).

Author information

Authors and Affiliations

  1. Department of Chemistry and Chemical Engineering, Heze University, Daxue Road, Heze, Shandong, 274015, People’s Republic of China

    Xinying Ma, Xueliang Wang & Meifeng Chen

  2. National Glycoengineering Research Center, Shandong University, Jinan, Shandong, 250012, People’s Republic of China

    Fuchuan Li

Authors
  1. Xinying Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xueliang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Meifeng Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Fuchuan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Xueliang Wang or Fuchuan Li.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ma, X., Wang, X., Chen, M. et al. Study on a new high affinity anti-glypicans-3 antibody in diagnosis of early hepatocellular carcinoma by differential pulse voltammetry. J Solid State Electrochem 21, 1631–1637 (2017). https://doi.org/10.1007/s10008-017-3535-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10008-017-3535-1

Keywords

Search

Navigation