Malaysian Journal of Analytical Sciences Vol 21 No 6 (2017): 1335 - 1341

DOI: 10.17576/mjas-2017-2106-15

 

 

 

DETECTION OF BIOMOLECULES BY A NOVEL FLUORESCENCE APTASENSOR BASED ON POLYDOPAMINE SPHERES

 

(Pengesanan Biomolekul oleh Aptasensor Pendarfluor Novel Berasaskan Sfera Polidopamina)

 

Haiyan Li¹ and Sam Fong Yau Li^{1, 2}*

 

¹Department of Chemistry,

National University of Singapore, 3 Science Drive 3, Singapore 117543, Singapore

²NUS Environmental Research Institute,

5A Engineering Drive 1, T-Lab Building, Singapore 117411, Singapore

 

*Corresponding author:  chmlifys@nus.edu.sg

 

 

Received: 7 November 2016; Accepted: 18 September 2017

 

 

Abstract

Aptamers are alternative bio-recognition elements to antibodies, with great advantages, such as high binding affinity and specificity, controllable structures. In this study, a novel fluorescent aptasensor for biomolecules detection has been designed by using aptamer as bio-recognition element and polydopamines as nanoquencher.  Bovine catalase protein (BCP) was used as target for assay validation. BCP aptamer was selected based on previous work in our group. Polydopamine spheres (250 – 300 nm) were synthesized by addition of dopamine hydrochloride into a mixture of tris-buffer and isopropyl alcohol with stirring for more than 2 days and size/morphology of spheres has been characterized. The quenching efficiency and sensing performance were examined. It was found that the quenching efficiency was good (> 90%) owing to the interaction between aptamer and polydopamines since FAM labeled ssDNA aptamer could be adsorbed onto polydopamines surface to form a complex, resulting in the quenching of fluorescence. Upon the introduction of BCP, the fluorescence could be retained. The fluorescence intensity increased with the increasing concentration of BCP with the detection limit of 10 nM. Polydopamines could be used as sensing platform in fast and simple way for biomolecules detection and have potential to be extended to other targets.

 

Keywords:  biosensors, aptamer, polydopamine, nanoquencher, protein

 

Abstrak

Aptamer adalah elemen alternatif pengecaman-bio terhadap antibodi, dengan kelebihan seperti ikatan keafinan yang kuat dan khusus, kebolehkawalan struktur. Dalam kajian ini, aptasensor pendarflour novel untuk pengesanan biomolekul telah di reka bentuk dengan mengunakan aptameter sebagai elemen pengecaman-bio dan polidopamina sebagai nanoquencher. Protein katalase Bovin (BCP) telah digunakan sebagai sasaran bagi ujian validasi. Aptameter BCP telah dipilih berdasarkan kerja terdahulu. Sfera polidopamina (250 – 300 nm) telah di sistesis melalui penambahan dopamina hidroklorik ke dalam campuran penimbal-tris dan alkohol isopropil dengan pengacauan melebihi 2 hari dan saiz/morfologi juga telah dicirikan. Keberkesanan pemelindapan dan prestasi sensor telah dikaji. Ia di dapati bahawa keberkesanan pemelindapan adalah baik (> 90%) terhadap interaksi antara aptameter dan polidopamina di ketahui bahawa aptamer ssDNA berlabel FAM berupaya menyerap ke dalam permukaan polidopamina untuk membentuk kompleks, menghasilkan pemelindapan pendarfluor. Apabila BCP didedahkan, sifat pendafluor dapat dikekalkan. Keamatan pendafluor meningkat seiring dengan peningkatan kepekatan BCP pada had pengesan ialah 10 nM. Polidopamina boleh digunakan sebagai platfom sensor yang pantas dan mudah bagi pegesanan biomolekul dan berpotensi untuk dikembangkan bagi sasaran lain.

 

Kata kunci:  sensor-bio, aptamer, polidopamina, nanoquencher, protein

 

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