Surface chemistry- and nanomaterials-enhanced high performance protein microarrays

There still remain great challenges to improve performance of protein microarrays in practical applications. Objectives of this project are to develop novel surface chemistry- and nanomaterials-based signal enhancement strategies, and to explore how they improve performance of protein microarrays. F...

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Main Author: Liu, Yingshuai
Other Authors: Li Changming
Format: Theses and Dissertations
Language:English
Published: 2011
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Online Access:https://hdl.handle.net/10356/45493
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-454932023-03-03T15:57:36Z Surface chemistry- and nanomaterials-enhanced high performance protein microarrays Liu, Yingshuai Li Changming School of Chemical and Biomedical Engineering Centre for Advanced Bionanosystems DRNTU::Science::Medicine::Biosensors DRNTU::Science::Chemistry::Physical chemistry::Surface chemistry There still remain great challenges to improve performance of protein microarrays in practical applications. Objectives of this project are to develop novel surface chemistry- and nanomaterials-based signal enhancement strategies, and to explore how they improve performance of protein microarrays. First, a poly(glycidyl methacrylate) brush is prepared and used as a supporting matrix, which provides high protein loading capacity, for improving performance of protein microarrays. Printing buffer, probe concentration, and immobilization kinetics are systematically investigated to elucidate fundamentals of protein attachment. The high performance is demonstrated by detections of hepatitis B virus surface antigen (HBsAg) in both phosphate buffered saline (PBS) and human serum. Second, a poly[glycidyl methacrylate-co-poly(ethylene glycol) methacrylate] (P(GMA-co-PEGMA)) brush is in situ synthesized on a poly(methyl methacrylate) (PMMA) substrate and silica nanoparticles (SNP) to synergistically amplify sensitivity of protein microarrays. In this design, the brush-coated SNP could carry a large number of reporter molecules at a single binding event to amplify detection signals, whereas the brush-coated PMMA offers high probe loading capacity and low nonspecific protein adsorption. The sensitivity of protein array is significantly improved. Third, a unique ZnO nanorods-coated substrate is developed to immobilize a large amount of probe molecules and also to directly amplify the microarray fluorescent signals. Two important cancer biomarkers, carcinoembryonic antigen (CEA) and α-fetoprotein (AFP), are analyzed in human serum with a detection limit of 1 pg mL-1, which is comparative to or better than that of ELISA. The advanced ZnO nanorods-based substrate is inexpensive, mass producible, and compatible with well-established microarray printing techniques, providing great potentials for developing economical and sensitive protein arrays. Fourth, a highly sensitive flow-through microarray immunoassay device is developed using a P(GMA-co-PEGMA) brush as a 3-D flexible matrix to achieve large probe loading capacity and a high bioactivity while allowing easy target access to probes in the brush. A limit of detection (LOD) of 1-10 pg mL-1 for detected targets is one to two orders better than those of reported flow-through immunoassays. DOCTOR OF PHILOSOPHY (SCBE) 2011-06-14T03:37:28Z 2011-06-14T03:37:28Z 2011 2011 Thesis Liu, Y. (2011). Surface chemistry- and nanomaterials-enhanced high performance protein microarrays. Doctoral thesis, Nanyang Technological University, Singapore. https://hdl.handle.net/10356/45493 10.32657/10356/45493 en 141 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Medicine::Biosensors
DRNTU::Science::Chemistry::Physical chemistry::Surface chemistry
spellingShingle DRNTU::Science::Medicine::Biosensors
DRNTU::Science::Chemistry::Physical chemistry::Surface chemistry
Liu, Yingshuai
Surface chemistry- and nanomaterials-enhanced high performance protein microarrays
description There still remain great challenges to improve performance of protein microarrays in practical applications. Objectives of this project are to develop novel surface chemistry- and nanomaterials-based signal enhancement strategies, and to explore how they improve performance of protein microarrays. First, a poly(glycidyl methacrylate) brush is prepared and used as a supporting matrix, which provides high protein loading capacity, for improving performance of protein microarrays. Printing buffer, probe concentration, and immobilization kinetics are systematically investigated to elucidate fundamentals of protein attachment. The high performance is demonstrated by detections of hepatitis B virus surface antigen (HBsAg) in both phosphate buffered saline (PBS) and human serum. Second, a poly[glycidyl methacrylate-co-poly(ethylene glycol) methacrylate] (P(GMA-co-PEGMA)) brush is in situ synthesized on a poly(methyl methacrylate) (PMMA) substrate and silica nanoparticles (SNP) to synergistically amplify sensitivity of protein microarrays. In this design, the brush-coated SNP could carry a large number of reporter molecules at a single binding event to amplify detection signals, whereas the brush-coated PMMA offers high probe loading capacity and low nonspecific protein adsorption. The sensitivity of protein array is significantly improved. Third, a unique ZnO nanorods-coated substrate is developed to immobilize a large amount of probe molecules and also to directly amplify the microarray fluorescent signals. Two important cancer biomarkers, carcinoembryonic antigen (CEA) and α-fetoprotein (AFP), are analyzed in human serum with a detection limit of 1 pg mL-1, which is comparative to or better than that of ELISA. The advanced ZnO nanorods-based substrate is inexpensive, mass producible, and compatible with well-established microarray printing techniques, providing great potentials for developing economical and sensitive protein arrays. Fourth, a highly sensitive flow-through microarray immunoassay device is developed using a P(GMA-co-PEGMA) brush as a 3-D flexible matrix to achieve large probe loading capacity and a high bioactivity while allowing easy target access to probes in the brush. A limit of detection (LOD) of 1-10 pg mL-1 for detected targets is one to two orders better than those of reported flow-through immunoassays.
author2 Li Changming
author_facet Li Changming
Liu, Yingshuai
format Theses and Dissertations
author Liu, Yingshuai
author_sort Liu, Yingshuai
title Surface chemistry- and nanomaterials-enhanced high performance protein microarrays
title_short Surface chemistry- and nanomaterials-enhanced high performance protein microarrays
title_full Surface chemistry- and nanomaterials-enhanced high performance protein microarrays
title_fullStr Surface chemistry- and nanomaterials-enhanced high performance protein microarrays
title_full_unstemmed Surface chemistry- and nanomaterials-enhanced high performance protein microarrays
title_sort surface chemistry- and nanomaterials-enhanced high performance protein microarrays
publishDate 2011
url https://hdl.handle.net/10356/45493
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