Surface Plasmon Resonance Analyzer

5 6 SPR System Based on Microfluidics Wide Dynamic Range Kinetic Analysis by Detection of Association /Dissociation of Bio-Molecules Measuring of Mass Change below ~10nM. The SPR signal originates in changes in the refractive index at the surface layer, caused by concentration changed on the sensor chip surface with adsorbed bio-molecules. The SPR is detected by measurement of the intensity of reflected light. From the SPR angle, a sharp decrease or dip in intensity can be measured. Any change in thickness or index of refraction of the material adsorbed onto gold surface results in a shift of the SPR angle. The SPR angle shift is used as a response unit to quantify binding molecules to the sensor surface. Principles of Photo diode Detector Polarized Light Analyte Solution of Protein Θ Glass (10mm) Coupling Prism Flow Cell Refractive Index Inject Ligand Inject Analyte Ligand Self assembled monolayer 50mm Gold Surface Cr (2nm) Au (50nm) SAM (~3nm) Substance Prism Time Surface refractive index is monitored over time as binding events occur.

SOFTWARE Measurement Mode Pre-scanning Mode (θ-r plot) : Incident angle vs Reflectance Pre-scanning mode is the method for measuring reflectance intensity as incident angle of the p-polarized light source and finding minimum reflectance angle. This mode is the angle resolution of 0.01 ~ 0.5 degree and measurement range within 30 ~ 80 degree. Fixed Angle Mode ( T-R plot) : Time vs Reflectance Fixed angle mode is the method for finding the maximum slope of the SPR curve and measuring the reflectivity with real time at this fixed angle. This process is performed in order to rapid monitoring at the minimum 0.2 second interval. As a measuring of change of reflectance with time function, this mode is displayed by Time vs Reflectance intensity change. Repeated Angle Mode ( T- Rθ plot): Time vs Resonance angle Repeated angle mode is the method for measuring the change of minimum resonance angle in the constant narrow angle range by repeatedly angle scanning and fitting. As a measuring of resonance angle shift with time function, this mode is displayed by Time vs Resonance angle change. Analysis Mode The interpretation of the measurement data and of the experimental result using the Simulation Mode, estimation of the experimental result, comparison with the result of theoretical and experimental.

7 8 SPECIFICATION DIMENSIONS Measurement Method Angle Scan Sensor & Sample Flow Sample Flow Channel 1 Sensing Point 1 Flow Tube 0.5mm(dia.) Light Source Semiconductor Laser(635nm, 2.5mW) Flow cell Material PEEK + Kalrez O-ring Sensor System Detector Incident Angle Range 1 Photodiode 30-80 degree Flow System Flow Tube Material Flow rate PEEK 5-100ul/min Incident Angle Change Mechanism Pulse Motor Sample Volume Sample Cell : 8.06 ul Injection Volume : < 100ul (changeable) Au Coated Substrate 18(W) 18(D) 10(H) mm Pump Method 1 Syringe Pump for Buffer and Sample Outer Dimensions Weight Power Supply Software(Window 98, 2000, XP) 600(D) 450(W) 270(H) mm 45 kg AC 220V, 60Hz SPR LAB Ver.2.X Temp. Control System Sample Injection Method Control Range Control Method Temp. Accuracy 2-Position Injection Valve(10port) 1st Loop for Sample, 2nd Loop for Air From 4 to 40 (5 /hr) Peltier Element ± 0.1 274.00 600.00 450.00 UNIT : mm

9 10 SPR Imaging System Based on Array type High Throughput Screening of Target Molecules. Rapid Monitoring of Biochip Ease to Use Powerful image Processing S/W With the advance of proteomics and the requirement of analysis of a variety of expressed proteins, rapid and high-throughput detection of the proteins is the hot issue. SPRi detects the specific binding of unlabeled bio-molecules to arrayed capture molecules attached to a modified gold thin film by measuring changes in the local index of refraction upon adsorption. It has been proven to be a valuable tool for investigating molecular interactions without the use of any fluorescent or radiochemical labels. SPR imaging using fast optical array detectors permits simultaneous measurements across an array of immobilized molecules. SPR imaging provides excellent spatial resolution at the same sensitivity as classical SPR. These features make SPR imaging a promising detection technology for biochips. Also, SPR imaging has applied to bioanalytical chemistry such as label-free investigation of enzyme substrate interactions, DNA hybridization, antibody antigen binding and protein DNA or protein protein. Principles of Parallel Beam Analyte Metal Layer Prism ΔR ra rr α Light Source CCD array Schematic of SPR imaging. A parallel beam is incident on a prism near the SPR resonance angle. The reflected light is detected by a CCD Array. Contrast in SPR image arises from variation in the reflected light intensity ΔR.

SOFTWARE Spot detection Viewer This viewer is display detection of target spot and information of spot, which is apply to setting range of detection spot and background. The detection method is automation and manual type. Line Profile Viewer It is the viewer that display line intensity profiling of X axis or Y- axis image using 2-Dimension image data 3 Dimension Viewer It is the viewer that display 3-Dimension image based on intensity of 2-Dimension image data, which is measuring or loading 3D Image Protein Concentration

11 12 SPECIFICATION DIMENSIONS Sensor System Measurement Method Sensing Point Light Source Detector Prism Material Incident Angle Change Mechanism Au Coated Substrate Intensity Interrogation Array Light Emitting Diode (760nm) 2D Charge Coupled Device (CCD) BK7, SF10, Fused Silica, BaCD4 or User Modify available Manual type 18(W) x 18(D) x 0.5(H) mm Outer Dimensions Weight Power Supply Software(Window 98, 2000, XP) 430(W) x 380(D) x 430(H) 30kg AC 220V, 60(Hz) MS window XP 422.50 433.00 370.19 UNIT : mm

GOLD CHIP The sensor chip is the signal transducer in real time SPR. The chip is a glass slide with a thin layer of gold deposited on one side. Gold is chosen for its combination of chemical inertness and good SPR response. The gold film on the sensor chip is covered with a covalently bound matrix to which bio-molecules may be immobilized using well defined chemistry. Types of Gold Chip Model Specipication G1 G2 G1 10 mm 12 mm 0.3 mm G2 18 mm 18 mm 0.3 mm G3 G4 G3 25Φ 1.0 mm G4 Block Prism 90 G5 G6 25mm 25mm 1.0mm, BK7 Silicone wafer(4") G5 G9 G7 SF10 Glass G8 Quartz cell G9 G10 Slide Glass Reaction Chamber G6 G11 Prism reference G12 Order Made

13 14 ACCESSORY Cleaning & Reaction Holder Constant Temperature Incubator Types Prism coupled chip holder (PCH5, PCH1) Slide chip holder (SCH) PCH5 PCH1 Power Supply Control Range Resolution Display Interface AC220V, 60Hz From 4 to 40 0.1 /min Character LCD Key pad type SCH1 The holders can be used SPR sensor chip cleaning and reaction such as SAM formation 136 222 360 UNIT : mm

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