| 000 | 09954nam a22002537a 4500 | ||
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| 008 | 211216b2012 |||a|||f mb|| 00| 0 eng d | ||
| 040 |
_aEG-CaNU _cEG-CaNU |
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| 041 | 0 |
_aeng _beng |
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| 082 | _a620 | ||
| 100 | 0 |
_aAyman Amin Abdel Hamid _9203 |
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| 245 | 1 |
_aNoble Metal Nanoparticles : _bGreen Synthesis and Application in KRAS Mutation Detection / _cAyman Amin Abdel Hamid |
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| 260 | _c2012 | ||
| 300 |
_a175 p. _bill. _c21 cm. |
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| 500 | _3Supervisor: Mohamed Abdel-Mottaleb | ||
| 502 | _aThesis (M.A.)—Nile University, Egypt, 2012 . | ||
| 504 | _a"Includes bibliographical references" | ||
| 505 | 0 | _aContents: ACKNOWLEDGEMENTS ................................................................................................ v LIST OF FIGURES ........................................................................................................... ix LIST OF TABLES ............................................................................................................ xv LIST OF EQUATIONS ................................................................................................... xvi LIST OF ACRONYMS .................................................................................................. xvii ABSTRACT ........................................................................................................................ 1 THESIS OBJECTIVES ...................................................................................................... 3 THESIS OUTLINE ............................................................................................................. 5 1 INTRODUCTION ...................................................................................................... 7 1.1 History of Nanoscience and Technology ................................................................... 7 1.2 Fabrication of Nanomaterials ..................................................................................... 8 1.3 Characterization of Nanomaterials ............................................................................. 8 1.4 Applications of Nanomaterials ................................................................................... 9 1.5 Green Synthesis of Metal Nanoparticles .................................................................. 10 1.5.1 Biosynthesis of Metal Nanoparticles Using Microorganisms .......................... 11 1.5.2 Biosynthesis of Metal Nanoparticles Using Whole Plants ............................... 13 1.5.3 Biosynthesis of Metal Nanoparticles Using Plant Extracts and Derivatives .... 15 1.6 Applications of Noble Metal Nanoparticles ............................................................. 26 1.6.1 Photothermal Applications................................................................................ 28 1.6.2 Drug Delivery ................................................................................................... 28 vii 1.6.3 Biosensing ......................................................................................................... 29 2 PHYTOSYNTHESIS OF Au, Ag AND Au-Ag BIMETALLIC NANOPARTICLES USING AQUEOUS EXTRACT OF SAGO PONDWEED (POTAMOGETON PECTINATUS L.) 46 2.1 Aim of Work ............................................................................................................ 46 2.2 Introduction .............................................................................................................. 46 2.3 Materials and Methods ............................................................................................. 49 2.3.1 Materials ........................................................................................................... 49 2.3.2 Preparation of Aqueous Extract of Potamogeton Pectinatus L. ....................... 49 2.3.3 Synthesis of Metal Nanoparticles ..................................................................... 49 2.3.4 Stability Study ................................................................................................... 50 2.3.5 Studying the Effect of Synthesis Variables on the Metal Nanoparticles .......... 50 2.3.6 Synthesis of Au-Ag Bimetallic Nanoparticles Using Different Au:Ag Molar Ratios 50 2.4 Characterization ....................................................................................................... 50 2.5 Results and Discussion ............................................................................................. 51 2.5.1 UV-vis Spectral Analysis .................................................................................. 52 2.5.2 Stability Study ................................................................................................... 53 2.5.3 Effect of Synthesis Variables on the Metal Nanoparticles ............................... 54 2.5.4 Effect of Variation of Au:Ag Molar Ratio on the Au-Ag Bimetallic Nanoparticles .................................................................................................................. 62 2.5.5 TEM Analysis ................................................................................................... 63 2.5.6 EDX Analysis ................................................................................................... 73 2.5.7 XRD Analysis ................................................................................................... 74 2.5.8 FTIR Analysis ................................................................................................... 78 2.5.9 Zeta Potential Analysis ..................................................................................... 81 3 KRAS MUTATION DETECTION USING MORPHOLINO-GOLD NANOPARTICLE PROBES ........................................................................................................................... 82 viii 3.1 Aim of Work ............................................................................................................ 82 3.2 Introduction .............................................................................................................. 82 3.2.1 Epidermal Growth Factor Receptor (EGFR) .................................................... 83 3.2.2 KRAS Gene Mutations as Biomarker for Response to Anti-EGFR Therapy ... 89 3.2.3 KRAS Mutation Detection Methods ................................................................. 94 3.2.4 DNA and Mismatch Detection Using MOR-Au NP Probes ........................... 101 3.3 Materials and Methods ........................................................................................... 102 3.3.1 Preparation of MOR-Au Nanoparticle Probes ................................................ 102 3.3.2 Screening of MOR Sequences ........................................................................ 103 3.3.3 A549 Cell Culture and Genomic DNA Extraction ......................................... 106 3.3.4 Target Amplification and Detection Using MOR-Au NP Probes .................. 106 3.3.5 Calculating the Selectivity of the MOR-Au NP Probes.................................. 110 3.4 Results and Discussion ........................................................................................... 110 3.4.1 MOR-Au Nanoparticle Probes ........................................................................ 110 3.4.2 Screening of MOR Sequences ........................................................................ 113 3.4.3 Using A549 Cancer Cell Line as a Model for Tumour Cells Harbouring KRAS G12S Mutation .............................................................................................................. 120 4 CONCLUSION ....................................................................................................... 128 REFERENCES ............................................................................................................... 129 | |
| 520 | 3 | _aAbstract: A green method was developed for the synthesis of metal nanoparticles. In addition, Au nanoparticles were used for the detection of KRAS gene mutations. Au, Ag and Au-Ag bimetallic nanoparticles were phytosynthesized using the aqueous extract of sago pondweed (Potamogeton pectinatus L.). The nanoparticles were fully characterized. The effect of the synthesis variables on the nanoparticles was investigated using UV-vis spectral analysis. Results showed that they were mostly spherical in shape, although other shapes as nanotriangles and hexagons were observed as well. Alloy-type Au-Ag nanoparticles could be specifically synthesized at pH 12. The nanoparticles were stable over 3 weeks. FTIR spectroscopy results indicate that the flavones and proteins present in the plant extract are responsible for the synthesis and stabilization of the nanoparticles. On the other hand, gold nanoparticles were used for the detection of KRAS mutations to predict the response to anti-EGFR antibody therapy in patients with metastatic colorectal carcinoma. Gold nanoparticles were labeled with morpholino (MOR) oligos and used as probes for the detection of mutations in the KRAS gene. Different MOR sequences were screened for the detection of G12S, G12V and G13D KRAS mutations using synthetic DNA. A549 cell line was used as a model for G12S KRAS mutation-harboring tumor cells. A549 genomic DNA was extracted and amplified. Results showed that the behavior of the MOR-Au NP probes vary according to length and sequence of the MOR oligos. The optimum MOR sequences for the detection of G12S, G12V and G13D KRAS mutations were selected. G12S KRAS mutation could be detected using our probe with a selectivity of 5% in wild-type genomic DNA. | |
| 546 | _aText in English, abstracts in English and Arabic | ||
| 650 | 4 |
_aNano- Science & Technology _9199 |
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| 655 | 7 |
_2NULIB _aDissertation, Academic _9187 |
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_aNano- Science & Technology _9199 |
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_2ddc _cTH |
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_c9188 _d9188 |
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