Using optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics
碩士 === 國立中正大學 === 化學暨生物化學研究所 === 99 === Measuring the kinetic constants of antigen-antibody interactions becomes critical in characterizing specific affinity and, hence, exploring the feasibility of using such interactions in clinical diagnosis. In this study, we used the optical waveguide-based par...
Main Authors: | , |
---|---|
Other Authors: | |
Format: | Others |
Language: | zh-TW |
Published: |
2011
|
Online Access: | http://ndltd.ncl.edu.tw/handle/05425759914430516160 |
id |
ndltd-TW-099CCU00065010 |
---|---|
record_format |
oai_dc |
spelling |
ndltd-TW-099CCU000650102015-10-28T04:06:37Z http://ndltd.ncl.edu.tw/handle/05425759914430516160 Using optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics 光波導式粒子電漿共振生化感測器對生物分子間交互作用動力學之研究 Wu, Chao-Ching 吳昭慶 碩士 國立中正大學 化學暨生物化學研究所 99 Measuring the kinetic constants of antigen-antibody interactions becomes critical in characterizing specific affinity and, hence, exploring the feasibility of using such interactions in clinical diagnosis. In this study, we used the optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics. For the first part, two configurations of biosensing chips were used. Chip1 has a one-dimensional surface grating structure with a period of 833 nm by stamping with UV glue on a glass slide and gold nanoparticles are modified on the opposite side of the glass slide. Chip2 has a grating structure with a period of 833 nm by stamping on a photopolymerizable sol-gel layer which is coated on a glass slide and gold nanoparticles are modified on the sol-gel layer. The planar waveguides for chip1 and chip2 are the glass slide and the photopolymerized sol-gel layer, respectively. Detection of the optical resonant reflective position from waveguide surface enables high sensitivity label-free biosensing. The light source in this setup was a diode-pumped laser (532nm). The sensor resolution achieved by the gold nanoparticles-modified biosensing chips was 2.8×10-5 RIU for chip1 and 1.1×10-4 RIU for chip2. The detection limit for anti-DNP antibody was determined to be 1.4×10-11 M for chip1 and 5.4×10-10 M for chip2. Unfortunately, even we could observe the signal-time sensorgram, we could not measure the kinetic constants because of the low S/N ratio achieved by these optical setups. For the second part, a simple and label-free biosensing method has been developed for the determination of anti-ovalbumin antibody (anti-OVA) and anti-Mouse IgG antibody and monitoring their biomolecular interactions by using the fiber-optic particle plasmon resonance (FOPPR) biosensor. The FOPPR sensor is based on gold nanoparticles-modified optical fiber where the gold nanoparticle surface has been modified by a mixed self-assembled monolayer for conjugation of receptor (ovalbumin or mouse IgG) to detect the corresponding analyte (anti-OVA or anti-mouse IgG). During the detection process, an analyte reacts with a receptor immobilized on the optical fiber and their interaction can be monitored in real-time. The detection limits for anti-OVA antibody and anti-mouse IgG were determined to be 2.4×10-9 M and 4.3×10-9 M, respectively. In addition, by assuming a Langmuir type of adsorption isotherm, the quantitative measurement of binding kinetics, including the association and dissociation rate constants, yields a ka of 5.5×103 M-1s-1 and a kd of 3.5×10-4 s-1 for OVA/anti-OVA, and a ka of 1.9×106 M-1s-1 and a kd of 1.7×10-2 s-1 for mouse IgG/anti-mouse IgG. We demonstrate that the FOPPR biosensor can study real-time biomolecular interactions. Chau, Lai-Kwan 周禮君 2011 學位論文 ; thesis 103 zh-TW |
collection |
NDLTD |
language |
zh-TW |
format |
Others
|
sources |
NDLTD |
description |
碩士 === 國立中正大學 === 化學暨生物化學研究所 === 99 === Measuring the kinetic constants of antigen-antibody interactions becomes critical in characterizing specific affinity and, hence, exploring the feasibility of using such interactions in clinical diagnosis. In this study, we used the optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics. For the first part, two configurations of biosensing chips were used. Chip1 has a one-dimensional surface grating structure with a period of 833 nm by stamping with UV glue on a glass slide and gold nanoparticles are modified on the opposite side of the glass slide. Chip2 has a grating structure with a period of 833 nm by stamping on a photopolymerizable sol-gel layer which is coated on a glass slide and gold nanoparticles are modified on the sol-gel layer. The planar waveguides for chip1 and chip2 are the glass slide and the photopolymerized sol-gel layer, respectively.
Detection of the optical resonant reflective position from waveguide surface enables high sensitivity label-free biosensing. The light source in this setup was a diode-pumped laser (532nm). The sensor resolution achieved by the gold nanoparticles-modified biosensing chips was 2.8×10-5 RIU for chip1 and 1.1×10-4 RIU for chip2. The detection limit for anti-DNP antibody was determined to be 1.4×10-11 M for chip1 and 5.4×10-10 M for chip2. Unfortunately, even we could observe the signal-time sensorgram, we could not measure the kinetic constants because of the low S/N ratio achieved by these optical setups.
For the second part, a simple and label-free biosensing method has been developed for the determination of anti-ovalbumin antibody (anti-OVA) and anti-Mouse IgG antibody and monitoring their biomolecular interactions by using the fiber-optic particle plasmon resonance (FOPPR) biosensor. The FOPPR sensor is based on gold nanoparticles-modified optical fiber where the gold nanoparticle surface has been modified by a mixed self-assembled monolayer for conjugation of receptor (ovalbumin or mouse IgG) to detect the corresponding analyte (anti-OVA or anti-mouse IgG). During the detection process, an analyte reacts with a receptor immobilized on the optical fiber and their interaction can be monitored in real-time. The detection limits for anti-OVA antibody and anti-mouse IgG were determined to be 2.4×10-9 M and 4.3×10-9 M, respectively. In addition, by assuming a Langmuir type of adsorption isotherm, the quantitative measurement of binding kinetics, including the association and dissociation rate constants, yields a ka of 5.5×103 M-1s-1 and a kd of 3.5×10-4 s-1 for OVA/anti-OVA, and a ka of 1.9×106 M-1s-1 and a kd of 1.7×10-2 s-1 for mouse IgG/anti-mouse IgG. We demonstrate that the FOPPR biosensor can study real-time biomolecular interactions.
|
author2 |
Chau, Lai-Kwan |
author_facet |
Chau, Lai-Kwan Wu, Chao-Ching 吳昭慶 |
author |
Wu, Chao-Ching 吳昭慶 |
spellingShingle |
Wu, Chao-Ching 吳昭慶 Using optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics |
author_sort |
Wu, Chao-Ching |
title |
Using optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics |
title_short |
Using optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics |
title_full |
Using optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics |
title_fullStr |
Using optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics |
title_full_unstemmed |
Using optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics |
title_sort |
using optical waveguide-based particle plasmon resonance biosensor for quantitative measurement of biomolecular interaction kinetics |
publishDate |
2011 |
url |
http://ndltd.ncl.edu.tw/handle/05425759914430516160 |
work_keys_str_mv |
AT wuchaoching usingopticalwaveguidebasedparticleplasmonresonancebiosensorforquantitativemeasurementofbiomolecularinteractionkinetics AT wúzhāoqìng usingopticalwaveguidebasedparticleplasmonresonancebiosensorforquantitativemeasurementofbiomolecularinteractionkinetics AT wuchaoching guāngbōdǎoshìlìzidiànjiānggòngzhènshēnghuàgǎncèqìduìshēngwùfēnzijiānjiāohùzuòyòngdònglìxuézhīyánjiū AT wúzhāoqìng guāngbōdǎoshìlìzidiànjiānggòngzhènshēnghuàgǎncèqìduìshēngwùfēnzijiānjiāohùzuòyòngdònglìxuézhīyánjiū |
_version_ |
1718111749021892608 |