Fabrication of an immunosensor for early and ultrasensitive determination of human tissue plasminogen activator (tPA) in myocardial infraction and breast cancer patients

  • Haidar Saify Nabiabad
  • Khosro Piri
  • Fatemeh Kafrashi
  • Abbas Afkhami
  • Tayyebeh Madrakian
Research Paper
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Abstract

Sensitive detection of biomarkers will mean accurate and early diagnosis of diseases. A tissue plasminogen activator (tPA) has a crucial role in many cardiovascular diseases and it is related to many processes such as angiogenesis in cancer cells. Therefore, sensitive determination of tPA is important in diagnosis and clinical research. tPA monoclonal antibody was covalently attached onto single-wall carbon nanotubes (SWCNTs) using diimide-activated imidation coupling. Functionalized SWCNTs were immobilized onto a glassy carbon electrode and the modification process was investigated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), SEM, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). Cyclic voltammograms (CVs) in a scan rate of 100 mVs−1 was studied and comparisons were made between the modified glassy carbon electrodes (immobilized with antibodies) as a working electrode before and after the formation of tPA-antibody complex. Results of the SDS-PAGE demonstrated that the antibody was covalently and site directly attached to the SWCNTs. The fabricated biosensor provided a good linear response range from 0.1 to 1.0 ng mL−1 with a low detection limit of 0.026 ng mL−1. The immunosensor showed selectivity, reproducibility, good sensitivity, and acceptable stability. Satisfactory results were observed for early and sensitive determination of tPA in human serum samples. For the first time, such specific biosensor is currently being fabricated for tPA in our laboratories and successfully could determine tPA in myocardial infraction and breast cancer patients.

Graphical abstract

Fabricated biosensor for determination of tPA

Keywords

Antibody functionalized SWCNTs Biosensor Cyclic voltammetry Tissue plasminogen activator (tPA) 

Abbreviations

CV

Cyclic voltammetry

Da

Dalton

EDC

1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide

DDW

Double-distilled water

GCE

Glassy carbon electrode

NHS

N-Hydroxy succinimide

EIS

Impedance spectroscopy

Ab

Monoclonal antibody

PBS

Phosphate buffer solution

MWCO

Por molecular weight cutoff

SEM

Scanning electron microscope

SWCNTs

Single-wall carbon nanotubes

SDS-PAGE

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis

tPA

Tissue plasminogen activator

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Notes

Compliance with ethical standards

Ethical approval for the study was granted by the Research Ethical Committee of the Bu-Ali Sina University (ethical code: IR.BASU.BIO.1392.26), and all patients gave written, informed consent.

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Haidar Saify Nabiabad
    • 1
  • Khosro Piri
    • 2
  • Fatemeh Kafrashi
    • 3
  • Abbas Afkhami
    • 3
  • Tayyebeh Madrakian
    • 3
  1. 1.Department of Medicinal Plant ProductionNahavand UniversityNahavandIran
  2. 2.Department of Biotechnology, College of AgricultureBu-Ali Sina UniversityHamadanIran
  3. 3.Department of Analytical Chemistry, Faculty of ChemistryBu-Ali Sina UniversityHamadanIran

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