Introduction to Biosensors

2007 Mattias Rudh www.realtimebiosensor.com

Definition of a biosensor
• A biosensor: • A device that uses specific biochemical reactions mediated by isolated enzymes, immunosystems, tissues, organelles or whole cells to detect chemical compounds usually by electrical, thermal or optical signals. Source: • PAC, 1992, 64, 148 (Glossary for chemists of terms used in biotechnology.)

Biosensor breakdown Analyte Response Analysis Signal Detection Sample handling/preparation .

vitamin. toxin.The Analyte What do you want to detect? Molecule Protein. peptide. sugar. metal ion Cholera toxin Glucose .

Sample handling How to do deliver the analyte to the sensitive region? •(Micro) fluidics •Concentration (increase/decrease) •Filtration/selection .

Example: Biosensor Application How to do deliver the analyte to the sensitive region? Collection wand .

Detection/Recognition Fab How do you specifically recognize the analyte? Active site Membrane receptors Competitive binding Fc Antibody Enzyme Cell Polymer/Hydrogel .

Conductivity) Electromechanical (QCM) Thermal Magnetic Pressure Often the detector is immobilized on a solid support/sensor . ELM.Signal How do you know there was a detection? Specific recognition? Common signaling principles Optical (SPR. IR) Electrical (Voltammetry. Potentiometry.

Avoiding false signals Specific recognition False specific recognition? Non specific signal .

fluorecent dye.Improving performance Secondary signal amplifier Highly specific detection Magnectic bead. enzyme etc Inert background vv v .

Regeneration or single use? Break binding Low and high pH buffers pH~1 and pH~13 vv v .

Data Analysis R t • Response variable (R) vs time(t): Example of response variables: Refractive index Potential Current Frequency Mass Pressure Temperature .

Baseline Should be stable when there is no binding Stable baseline Drift baseline t Quantifying Noise Root mean square (RMS) of a sample of data points for a given time Quantifying Drift Shift in the baseline (RMS) shown as response units per time t .

Sensitivity Signal-to-noise ratio Per time unit t Spikes Rapid (1 datapoint!) shift in signal Baseline shift Rapid (1 datapoint!) shift in baseline (offset) t t .

Common signal error sources • • • • • • Inhomogenous sample Bubbles/flow artifacts Temperature Electromagnetic interferance Electronic unstability Unstable chip/detection layer .

Improved sensitivity Active sensor detects the analyte Reference sensor Coated with inert material does not detect the analyte R2 R1 Output signal R=R1-R2 or R=R1/R2 The reference is exposed to the same kind of disturbances as the active sensor. These effects are cancelled out by taking the difference between the two sensors Sample R1 t R2 t R t .

Signal interpretation • Visual (example pregnancy test) • Automatic (Software) • Manual (Research Biosensor) .

Kinetic evaluation • Binding / no binding • Affinity (Ka / Kd and k_on and k_off) .

Example of biosensors Pregnancy test Detects the hCG protein in urine. . Interpretation and data analysis performed by the user Glucos monitoring device (for diabetes patients) Monitors the glucose level in the blood. Interpretation and data analysis performed by a microprocessor.

.Example of biosensors Infectous disease biosensor from RBS Data analysis and interpretatoin performed by a microprocessor Old time coal miners’ biosensor Data analysis and interpretation performed by the coal miner.

com .realtimebiosensor. good tool for development of specific biosensors For a comprehensive list of research biosensor suppliers see: www.Research Biosensors Biacore Biosensor platform General and flexible.

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