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Development of Nanoprobes for the Biological Imaging Using Positive Charged Quantum Dots and Upconversion Nanoparticles

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dc.contributor.author이, 정한-
dc.date.accessioned2011-11-03T05:43:38Z-
dc.date.available2011-11-03T05:43:38Z-
dc.date.issued2011-
dc.identifier.urihttp://repository.ajou.ac.kr/handle/201003/4348-
dc.description.abstractNanoparticles (NPs) have unique physicochemical properties due to the quantum confinement effect. Semiconductor quantum dots (QDs) or upconversion nanoparticles (UCNPs) exhibit interesting properties such as high photoluminescence, photostability, dual magnetic/optical modality. In this study, biocompatible and stable NPs were developed through the various surface modifications. NPs were functionalized by its conjugation with various molecules including antibody for detection of kappa opioid receptor, small peptide for detection of integrin ανβ3 and oligonucleotides for the detection of influenza virus. These functionalized NPs were useful to visualize the endocytosis of kappa opioid receptor in live cell, to acquire in vivo imaging of tumor expressing integrin ανβ3 in mice, and to detect sequence-specific oligonucleotides. These results show the potential of these nanoprobes to apply for the drug screening and in vivo multiplex imaging.-
dc.description.abstractQuantum dot 혹은 upconversion nanoparticle과 같은 나노물질들은 그들의 고유한 물리 화학적 특성 (광안정성, 높은 양자수율, 입자크기 조절에 따른 형광파장의 변화, 광학-자기적 특성 등)으로 인해, 생물학 분야에서 기존에 사용되고 있던 유기 형광체를 대체할 물질로 각광받고 있다. 본 연구에서는 다양한 표면개질을 통해 친수화시킨 나노입자를 이용하여, kappa opioid receptor를 표적으로 한 항체, integrin ανβ3 targeting을 위한 peptide 및 influenza virus 검출을 위한 DNA 등과 같은 다양한 biomolecule을 결합시킨 후, 이를 이용하여 살아있는 세포에서 kappa opioid receptor의 endocytosis 과정을 시각화, 동물모델에서 integrin ανβ3 를 발현하는 tumor의 이미징, .혹은 sequence-specific 표적 DNA의 검출 가능성을 보여주었다. 이 결과들을 통해 신약 개발에 필요한 screening 분야로의 응용 가능성과 in vivo multiplex imaging을 위한 탐침제로써의 활용 가능성을 보여주었다.-
dc.language.isoen-
dc.titleDevelopment of Nanoprobes for the Biological Imaging Using Positive Charged Quantum Dots and Upconversion Nanoparticles-
dc.title.alternative양전하를 띠는 양자점 및 업컨버젼 나노입자를 이용한 생물학적 이미징을 위한 탐침제 개발-
dc.typeThesis-
dc.identifier.urlhttp://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000011387-
dc.subject.keywordkappa opioid receptor-
dc.subject.keywordintegrin ανβ3-
dc.subject.keywordinfluenza virus-
dc.subject.keywordendocytosis-
dc.subject.keywordtumor imaging-
dc.subject.keyworddurg screening-
dc.subject.keywordin vivo multiplex imaging-
dc.subject.keywordQD-
dc.subject.keywordUCNP-
dc.description.tableOfContentsABSTRACT ⅰ

TABLE OF CONTENTS ⅱ

LIST OF FIGURES ⅴ

LIST OF TABLES ⅸ

Ⅰ. INTRODUCTION 1

A. Surface modification of Quantum Dots (QDs) 2

1. Pegylated amine functionalized QD 3

2. QD-DNA sensor based on FRET 3

B. Application of QD for the target protein detection 5

1. QD labeled kappa opioid receptor and drug induced endocytosis 5

2. Application of nanoprobes to drug screening method 7

C. Development of multimodal imaging nanoprobe 8

1. Development of nanoprobes for in vivo tumor targeting 8

2. Multi-imaging modality of UCNP 8

Ⅱ. MATERIALS AND METHODS 9

A. MATERIALS 9

1. CdSe/ZnS QDs synthesis and surface modification with various ligands and polymer 9

2. Positively charged QD and oligonucleotide complex as a target DNA sensing material 9

3. Anti-HA antibody conjugated QDs and κ-OR targeting 11

4. Cyclo RGD conjugated UCNP and integrin ανβ3 binding experiment 11

B. METHODS 12

1. Nanoparticle synthesis and surface modification 12

2. Preparation of QD-antibody conjugates and GPCR targeting 17

3. UCNP-cRGD peptide conjugates and in vivo tumor targeting 19

4. General characterization methods 23

Ⅲ. RESULTS 25

A. Nanoparticle synthesis and surface modification 25

1. Synthesis of CdSe/ZnS QDs and polymer coating 25

2. Characterization of DEDEA ligand exchanged QD and pegylation 26

3. Complex formation of cationic QD with single-stranded oligonucleotides 31

4. Preparation of FRET-based QD-DNA probes 34

5. FRET efficiency of the QD-DNA complexes 35

6. Detection of target oligonucleotides 36

B. Nanoparticles functionalized with biomolecules for in vitro protein imaging 39

1. Amine functionalization of QD 39

2. Preparation of QD-anti-HA antibody conjugates 40

3. Specific binding of QD-anti-HA antibody conjugates to κ-OR 42

4. Endocytosis of QD labeled κ-opioid receptor by agonist 43

5. Real time tracking of κ-opioid receptor internalization 44

6. Endocytosis of κ-OR dependent on its specific agonist 45

7. Endocytosis pathway of κ-OR 46

8. The concentration-dependent effect of κ-OR agonist is visualized using QD-anti-HA antibody conjugates 47

C. Nanomaterials for deep tissue and in vivo imaging study 48

1. Cyclo RGD peptide-conjugated UCNPs 48

2. UCNP-(cRGD)2 bound to U87MG cells highly expressing integrin ανβ3 49

3. T1 weighted MR/micro PET images of mouse bearing U87MG glioma 51

4. Visualization of UCNP in tumor tissue 55

Ⅳ. DISCUSSION 57

A. Pegylated DEDEA-QD and application to production of the QD-DNA probe 57

B. QD-antibody conjugates for κ-ORs targeting and application to drug screening method 58

C. Synthesis of UCNP-(cRGD)2 conjugates and in vivo tumor imaging 59

Ⅴ. CONCLUSION 61

REFERENCES 63

국문요약 72
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dc.description.degreeDoctor-
dc.contributor.affiliatedAuthor이, 정한-
dc.date.awarded2011-
dc.type.localTheses-
dc.citation.date2011-
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