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

The Analyte
What do you want to detect? Molecule Protein, toxin, peptide, vitamin, 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

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

Avoiding false signals

Specific recognition False specific recognition?

Non specific signal

Improving performance
Secondary signal amplifier

Highly specific detection

Magnectic bead, fluorecent dye, 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

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

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)

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.

Research Biosensors
Biacore Biosensor platform General and flexible, good tool for development of specific biosensors

For a comprehensive list of research biosensor suppliers see: www.realtimebiosensor.com