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Boaz Vilozny October 4, 2011

BME 140 Bioengineering

What is a sensor?

A sensor is any device that converts a measurement into a signal

What is a biosensor?

A sensor in which the receptor is a biomolecule A sensor in which the analyte is a biomolecule A sensor in which the signal is produced by biological interactions

Optical Biosensors

Indicators and immobilized sensors

Optical biosensor: DNA microarray

What is the receptor? What is the analyte? How is the signal transduced? Is this a true sensor?

Fluorescent biosensor: Cameleon

What is the receptor?

What is the reporter? How is the signal transduced? How is sensor delivered to cells?

Electrical Biosensors

What is the electrical signal?

Potentiometry Amperometry Voltammetry Conductance, etc.

No optical components Miniaturizable Sensitivity increases with nanofabrication

Receptors for electrical biosensors

Enzymes Antibodies DNA/RNA

Electrical biosensor design

Sandwich Immunoassay

Urban, 2009 Measurement Science and Technology

Bioconjugation and biosensor fabrication

How is bioconjugation used? Requirements of bioconjugation




Non-chemical affinity methods


acid hybridization Protein/substrate interactions

Bioconjugation: chemical crosslinking

Why a chemical bond? Chemistry for special surfaces




polymers Charged surfaces

Optical immunoassay on a chip

Surface treatment Amino-Silanization Crosslinker Capture antibody Protein-blocking Analyte (IgG) Labeled antibody Optical measurement

Raj, J. et al, Biosens. Bioelectron. 2009

Voltammetric aptamer-biosensor

Gold surface on glassy carbon electrode

Doxorubicin as indicator: redox active, dsRNA intercalator

Why not a sandwich assay? How to achieve selectivity? How is signal generated?
Zhao, G-C. et al, Electrochem. Commun. 2010

Putting it all together: the biosensor as a self-contained diagnostics lab

Chin, C.D.; Sia, S.K. et al. Nature Medicine, 2011

The problem: low-cost diagnostics

Resource-limited settings

of medical facilities Lack of doctors Lack of money

Health epidemic Diagnostics can make a difference


siphylis treatable; avoids transmission to newborns

Why bioengineering?

Microfluidics Biochemistry Nanotechnology Medicine Engineering

The challenge of integration

Innovations: 1) Chip manufacturing (injection-molded plastic) 2) Reagent delivery (air spacing) 3) Signal amplification (silver reduction)

Chin, C.D.; Sia, S.K. et al. Nature Medicine, 2011

Antibody detection: ELISA

Immobilized antigen Target antibody

Gold-labeled secondary antibody

Signal amplification

Liquid handling

Does it work?

70 specimens tested 15 minutes, 1L blood One false positive (96% specificity)

Chin, C.D.; Sia, S.K. et al. Nature Medicine, 2011

Biosensing with nanopipettes

Nanopipettes as sensors

Fabrication Ionic current Current rectification

Ion current through nanopipettes

An electrical DNA nanosensor

Detected 15-base oligomers Signal detected by current rectification

Fu, Baker, et al Chem. Commun. 2009

A renewable nanosensor

Reporter dye trapped in pipette tip Fluorescence increases in presence of sodium Imaged with confocal microscope

Piper, Klenerman et al, J. Am. Chem. Soc. 2006

Label-free immunosensing with nanopipettes (STING)

Art by David Liao

Selective sensing with STING

Signal Transduction by Ion Nano-Gating

Binding to receptors modulates current through the conical nanopore

Electrical antibody recognition

1.1 1.0


I, normalized

0.9 0.8 -0.8 -0.9 -1.0 -1.1 -20 -10 0 10 20

Anti-VEGF Anti-ferritin

Time [min]

Umehara, Pourmand et al, Proc. Nat. Acad. Sci. 2009

Calcium ions: a good target for biosensing

Major affector of cellular events Indicator of cytotoxycity/apoptosis Single-cell arrays used to monitor cellular events by monitoring cytosolic [Ca2+] 1,2

1. Xu, et al. Anal. Biochem. 2010

2. Li et al. Integr. Biol. 2009

Li, 2009

A receptor for calcium: Calmodulin

Quartz surface functionalization

Change bath

Ca2+ (0.1 mM)

buffer (pH 7)


Calcium sensor is reversible






And selective


Nanopipettes modified with Calmodulin as a recepter Reversible calcium signal Selective over magnesium Detection limit 2x10-5 M Ca2+. Further work: improve surface chemistry, sensitivity, shelf life. Test with biological systems


Biosensors: form and function Bioconjugation: making biomolecules behave Biosensor case studies

sandwich immunoassay Electrical aptamer sensor mCHIP immunoassay STING bioelectrical sensor