2019. 7. 24 高階醫藥研發轉體與研究平台實作研習營
TRANSCRIPT
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2019. 7. 24 高階醫藥研發轉體與研究平台實作研習營
局部區域表面電漿共振LSPR (Localized Surface Plasmon Resonance)
Speaker : 林俐吟 & 游季禹
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1.什麼是表面電漿共振SPR (Surface Plasmon Resonance)?
2. SPR的重要性
3. 新一代的局部區域表面電漿共振LSPR (Localized Surface Plasmon Resonance)
4. 應用實例分享
5. LSPR操作流程
6. LSPR上機實作
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Surface Plasmon Resonance
Label-Free Interaction Analysis
1.什麼是表面電漿共振SPR (Surface Plasmon Resonance)?
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General Overview of an SPR Experiment
Time
Res
po
nse
1. Sensor Chemistry
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General Overview of an SPR Experiment
2. Immobilize ligand
Time
Res
po
nse
1. Sensor Chemistry
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General Overview of an SPR Experiment
2. Immobilize ligand 3. Introduce analyte at multiple concentrations
Time
Res
po
nse
1. Sensor Chemistry
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Binding Kinetics: Analyte Binding Curve
ka kd
A + B ⇌ AB𝑘𝑎
𝑘𝑑
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Binding Kinetics: Analyte Binding Curve
kd
𝑘𝑎
𝑘𝑑
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Binding Kinetics: Analyte Binding Curve
𝑘𝑎
𝑘𝑑
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Binding Kinetics: Analyte Binding Curve
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Binding Kinetics: Analyte Binding Curve
• Repeat 3 to 5 different concentrations of analyte
REPEAT
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Sensor Chips
Covalent Coupling Capture Coupling Lipid Capture
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Sensor Chips
Covalent Coupling
• Carboxyl
• Amine
• Thiol
Capture Coupling
• Streptavidin/Biotin
• NTA
• GST
• Protein A
• Liposome (LIP)
• Hydrophobic (MEM)
Lipid Capture
High sensor repeatability
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Applications
• Protein-Protein• Protein-Antibody• Protein-Nucleic Acid• Protein-Lipid
Materials scienceDetectionQuantificationNanomaterial characterization
• Protein-Carbohydrates• Protein-Small Molecule• Protein-Peptide• Protein-Aptamer
Binding Kinetics/Equilibrium Constants:• DNA, RNA• Virus particles• Cells & bacteria• Nanoparticles• Polymers
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kon
koff
KD
Yes/No Binding
SPR uncovers what’s underneath the surface16
Pull down
EMSA
2. SPR的重要性
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Affinity Doesn’t Tell the Whole Story
Different interactions can have the same affinity.
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Scientific publications involving Surface Plasmon Resonance (SPR) have increased drastically over the years.
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Traditional SPR Measurement Technique
Traditional SPR
Sensor Surface:• Thin film of gold• Surface plasmon resonance
Detection:• Change in refractive index• Reflectance angle
3. 新一代的局部區域表面電漿共振LSPR (Localized Surface Plasmon Resonance)
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LSPR Measurement Technique (OpenSPR)
Sensor Surface:• Nano-structured gold• Localized Surface plasmon resonance
Detection:• Change in refractive index• Absorbance (change in peak position)
LSPR
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LSPR SPR
• Bench Top instrument
• User friendly
• Save time
• Low-Maintenance • High throughput
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抗憂鬱藥的旅程
4. 應用實例分享
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1. 放置實驗所需之buffer以及DI water 2. 開啟儀器電源
3. 開啟電腦桌面之軟體
LSPR操作流程
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`
此步驟非必要,除非:1. 距離上次波長超過一個月2. 換LED光源時3. 訊號不足時
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Front sensorsurface
Cut corner of the glass positioned in the bottom right
STEP 5
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STEP 6
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Thank you for your attention!