Paper-based immunosensor with signal amplification by enzyme-labeled anti-p16INK4a multifunctionalized gold nanoparticles for cervical cancer screening

Ruchuon Yokchom; Somsak Laiwejpithaya; Weerakanya Maneeprakorn; Satita Tapaneeyakorn; Jundee Rabablert; Tararaj Dharakul

doi:10.1016/j.nano.2018.01.016

Paper-based immunosensor with signal amplification by enzyme-labeled anti-p16^INK4a multifunctionalized gold nanoparticles for cervical cancer screening

Nanomedicine. 2018 Apr;14(3):1051-1058. doi: 10.1016/j.nano.2018.01.016. Epub 2018 Feb 3.

Authors

Ruchuon Yokchom¹, Somsak Laiwejpithaya², Weerakanya Maneeprakorn³, Satita Tapaneeyakorn³, Jundee Rabablert⁴, Tararaj Dharakul⁵

Affiliations

¹ Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
² Department of Obstetrics & Gynaecology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
³ National Nanotechnology Center, National Science and Technology Development Agency, Thailand Science Park, Pathum Thani, Thailand.
⁴ Department of Biology, Faculty of Science, Silpakorn University, Nakorn Pathom, Thailand.
⁵ Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand. Electronic address: tararaj.dha@mahidol.ac.th.

PMID: 29407199
DOI: 10.1016/j.nano.2018.01.016

Abstract

The aim of this study was to develop a paper-based immunosensor for cervical cancer screening, with signal amplification by multifunctionalized gold nanoparticles (AuNPs). The AuNPs were functionalized with a highly specific antibody to the p16^INK4a cancer biomarker. The signal was amplified using a combination of the peroxidase activity of horseradish peroxidase (HRP) enzyme-antibody conjugate and the peroxidase-like activity of the AuNPs. The immune complex of p16^INK4a protein and multifunctionalized AuNPs was deposited on the nitrocellulose membrane, and a positive result was generated by catalytic oxidation of peroxidase enzyme substrate 3,3',5,5'-Tetramethylbenzidine (TMB). The entire reaction occurred on the membrane within 30 min. Evaluation in clinical samples revealed 85.2% accuracy with a kappa coefficient of 0.69. This proof of concept study demonstrates the successful development of a highly accurate, paper-based immunosensor that is easy to interpret using the naked eye and that is suitable for cervical cancer screening in low-resource settings.

Keywords: Cervical cancer screening; Multifunctionalized gold nanoparticles; Paper-based immunosensor; Signal amplification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies, Monoclonal / administration & dosage
Antibodies, Monoclonal / immunology
Benzidines / metabolism
Biosensing Techniques / methods*
Carcinoma, Squamous Cell / diagnosis
Carcinoma, Squamous Cell / immunology
Carcinoma, Squamous Cell / metabolism
Case-Control Studies
Cyclin-Dependent Kinase Inhibitor p16 / immunology*
Early Detection of Cancer / methods*
Female
Gold / chemistry*
Horseradish Peroxidase / chemistry*
Horseradish Peroxidase / metabolism
Humans
Immunoassay
Metal Nanoparticles / administration & dosage*
Metal Nanoparticles / chemistry
Paper*
Precancerous Conditions / diagnosis
Precancerous Conditions / immunology
Precancerous Conditions / metabolism
Uterine Cervical Neoplasms / diagnosis*
Uterine Cervical Neoplasms / immunology
Uterine Cervical Neoplasms / metabolism

Substances

Antibodies, Monoclonal
Benzidines
CDKN2A protein, human
Cyclin-Dependent Kinase Inhibitor p16
3,3',5,5'-tetramethylbenzidine
Gold
Horseradish Peroxidase