Use of liposomal amplifiers in total internal reflection fluorescence fiber-optic biosensors for protein detection

Ying-Feng Chang; Chen Fu; Yi-Ting Chen; Amily Fang-Ju Jou; Chii-Chang Chen; Chien Chou; Ja-An Annie Ho

doi:10.1016/j.bios.2015.10.012

Use of liposomal amplifiers in total internal reflection fluorescence fiber-optic biosensors for protein detection

Biosens Bioelectron. 2016 Mar 15:77:1201-7. doi: 10.1016/j.bios.2015.10.012. Epub 2015 Oct 8.

Authors

Ying-Feng Chang¹, Chen Fu², Yi-Ting Chen², Amily Fang-Ju Jou¹, Chii-Chang Chen³, Chien Chou⁴, Ja-An Annie Ho⁵

Affiliations

¹ BioAnalytical Chemistry and Nanobiomedicine Laboratory, Department of Biochemical Science and Technology, National Taiwan University, Taipei 10617, Taiwan.
² Department of Chemistry, National Tsing Hua University, Hsinchu 30013, Taiwan.
³ Department of Optics and Photonics, National Central University, Taoyuan 32001, Taiwan.
⁴ Graduate Institute of Electro-Optical Engineering, Chang Gung University, Taoyuan 33302, Taiwan; Healthy Aging Research Center, Chang Gung University, Taoyuan 33302, Taiwan.
⁵ BioAnalytical Chemistry and Nanobiomedicine Laboratory, Department of Biochemical Science and Technology, National Taiwan University, Taipei 10617, Taiwan; Department of Chemistry, National Tsing Hua University, Hsinchu 30013, Taiwan. Electronic address: jaho@ntu.edu.tw.

PMID: 26595485
DOI: 10.1016/j.bios.2015.10.012

Abstract

Evanescent-wave excited fluorescence technology has been demonstrated to enhance sensitivity and reduce matrix effects, making it suitable for biosensor development. In this study, we developed a liposome-based, total internal reflection fluorescence, fiber-optic biosensor (TIRF-FOB) for protein detection, which integrates a liposomal amplifier and sandwich immunoassay format with TIRF-FOB. In addition, the antibody-tagged and fluorophore-entrapped liposomes for heterogeneous detection of target molecules were designed and synthesized. This biosensor successfully detected the target protein (model analyzed here is IgG) with a limit of detection (LOD) of 2.0 attomoles for the target protein (equivalent to 2.0 pg/mL of protein presented in 150 μL of sample solution). The features of this ultra-sensitive liposomal TIRF-FOB are (i) fluorescence is excited via evanescent waves and amplified via liposomes; (ii) the use of two polyclonal antibodies in the sandwich assay format increases the specificity and lowers the cost of our assay. Based on the exceptional detection sensitivity and cost-effectiveness, we believe that the proposed biosensor has great potential as a practical, clinical diagnostic tool in the near future.

Keywords: Evanescent wave-excited fluorescence; Fiber-optic biosensors; Liposomes; Protein detection.

Publication types

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

MeSH terms

Biosensing Techniques / instrumentation*
Equipment Design
Equipment Failure Analysis
Fiber Optic Technology / instrumentation*
Immunoassay / instrumentation
Immunoglobulin G / analysis*
Liposomes / chemistry*
Protein Array Analysis / instrumentation*
Spectrometry, Fluorescence / instrumentation*

Substances

Immunoglobulin G
Liposomes