Enzyme-free impedimetric biosensor-based molecularly imprinted polymer for selective determination of L-hydroxyproline

Whitchuta Jesadabundit; Sakda Jampasa; Kanitha Patarakul; Weena Siangproh; Orawon Chailapakul

doi:10.1016/j.bios.2021.113387

Enzyme-free impedimetric biosensor-based molecularly imprinted polymer for selective determination of L-hydroxyproline

Biosens Bioelectron. 2021 Nov 1:191:113387. doi: 10.1016/j.bios.2021.113387. Epub 2021 Jun 3.

Authors

Whitchuta Jesadabundit¹, Sakda Jampasa², Kanitha Patarakul³, Weena Siangproh⁴, Orawon Chailapakul⁵

Affiliations

¹ Electrochemistry and Optical Spectroscopy Center of Excellence (EOSCE), Department of Chemistry, Faculty of Science, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand.
² Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Patumwan, Bangkok, 10330, Thailand.
³ Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand.
⁴ Department of Chemistry, Faculty of Science, Srinakharinwirot University, Sukhumvit 23, Wattana, Bangkok, 10110, Thailand. Electronic address: weena@g.swu.ac.th.
⁵ Electrochemistry and Optical Spectroscopy Center of Excellence (EOSCE), Department of Chemistry, Faculty of Science, Chulalongkorn University, Pathumwan, Bangkok, 10330, Thailand; National Center of Excellent of Petroleum, Petrochemicals, and Advanced Materials, Chulalongkorn University, Phayathai Road, Pathumwan, Bangkok, 10330, Thailand. Electronic address: corawon@chula.ac.th.

PMID: 34146970
DOI: 10.1016/j.bios.2021.113387

Abstract

This study first reported enzyme-free impedimetric biosensor-based molecularly imprinted polymers for selective and sensitive determination of L-hydroxyproline (L-hyp), a biomarker for the early diagnosis of bone diseases. In recent study, utilizing a single 3-aminophenylboronic acid (3-APBA) to create imprinted surfaces could result in a strong interaction and difficulty in removal of a template molecule. Hence, a mixture of monomer solution containing 3-APBA and o-phenylenediamine (OPD) in the presence of the L-hyp molecule was co-electropolymerized onto the screen-printed electrode using cyclic voltammetry (CV) to eradicate this mentioned limitation. The detection principle of this sensor is relied on alteration of mediator's charge transfer resistance (R_ct) that could be obstructed by L-hyp occupied in imprinted surface. The successfully fabricated biosensor was explored by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), and confocal scanning microscopy. Furthermore, the effect of polymer composition on the R_ct response was systematically investigated. The result exhibited that the mixture of monomers could provide the highest change of R_ct due to high selectivity from esterification of 3-APBA and from hydrogen bond of OPD surrounding the template. The sensor showed a significant increase in R_ct in the presence of L-hyp, whereas no observable resistance change was detected in the absence thereof. The calibration curve was obtained in the range from 0.4 to 25 μg mL^-1 with limits of detection (3SD_blank/Slope) and quantification (10SD_blank/Slope) of 0.13 and 0.42 μg mL^-1, respectively. This biosensor exhibited high selectivity and sensitivity and was successfully applied to determine L-hyp in human serum samples with satisfactory results.

Keywords: Bone diseases; Electrochemical impedance spectroscopy; L-hydroxyproline; Molecularly imprinted polymer; Non-enzymatic sensor.

MeSH terms

Biosensing Techniques*
Electrochemical Techniques
Electrodes
Humans
Hydroxyproline
Limit of Detection
Molecular Imprinting*
Molecularly Imprinted Polymers

Substances

Molecularly Imprinted Polymers
Hydroxyproline