BioDevice Technology, Ltd. BDT is a company born from Japan Advanced Institute of Science and Technology (JAIST) and was established in July 2003 by Professor Eiichi Tamiya and five faculties.

High Sensitive Immunochromatography PCT/JP2006/323617

DEP-Chip series for electrochemical biosensors

Electrochemical DNA sensors (JP3642574)

Gold-Linked Electrochemical Immuno Assay GLEIA, PCT/JP2007/56992)

Our Objects of Establishment Establish products as fruits of research in

universities. Back up of making collaboration academic

researchers with companies. Build innovation by collaboration among

academic, companies and government

R & D of Biosensors

Disposable Type Screen Printed Electrodes

DEP-Chip Series

SP-N, SP-P (carbon)

SR-N (gold)

Square working electrode

PCR-P01

EP-N (carbon) EP-PP (carbon, WE with ring and dam) ER-N (gold)

Round working electrode

One drop test Adapted size for PCR tube mass production/disposal/low cost Possible customized fabrication

Pathogenic bacteria/virus Salomonella, Ecoli O-157, Hepatis virus B, Influ virus Food safety GMO, Meat quality SNPs ApoE, ADH

Printable electrodes based biosensors

Gene sensing

Enzyme activity sensing

Gold Linked Electrochemical Immuno Assay : GLEIA hCG hormon : 36 pg/mL (612 fM) insulin : 125pg/mL

Antibody sensing

Acetylcholine esterase inhibition Organophosphate pesticide : 0.01 ppm HIV protease inhibition

Original principle of DNA sensor Japanese patent

A B

(Low concentration of DNA

or Before amplification)

(High concentration of DNA

or After amplification)

00.2

0.4

0.6

0.8

1

100 200 300 400 500 600 700

0

0.5

1

1.5

2

2.5

100 200 300 400 500 600 700

a

b

c

d

e

0

0.5

1

1.5

2

2.5

200 300 400 500 600 700 800

a

b

c

M a b c d e

M a b c

M a b c

0

0.4

0.8

1.2

1.6

2

200 300 400 500 600 700 800

a

b

c

M a b

Salmonella invA gene (PCR)

O157 gene (LAMP)

negative

positive

Genetically-modified corn (LAMP)

Hepatitis B virus gene from

blood sample (Nested PCR)

negative

positive

negative

negative

positive positive

Examples of various DNA targets detected

Analyst,136, 2064 (2011)

Rapid detection for screening of influenza A virus:

microfluidic RT-PCR chip and electrochemical DNA sensor

Analyst, 2011, Advance ArticleDOI: 10.1039/C1AN15638A

Semi-real time electrochemical monitoring for influenza virus RNA by

RT-LAMP using USB powered portable potentiostat

BDT miniSTAT BDT DEP chip

Gold nanoparticle redox signal enhancement for detection of antigens

After the recognition reaction between the surface-immobilized primary antibody and hCG, the captured antigen

was sandwiched with a secondary antibody that was labeled with gold-nanoparticles. Then the amount of

gold nanoparticles was determined by its redox signal.

The primary antibody was Immobilized directly on the working electrode, and a sandwich-type immuno-reaction was performed

A high potential was applied In HCl for the oxidation of gold nanoparticles

DPV* : differential pulse voltanmetry

The voltammetric measurement was performed.

References Direct fabrication of catalytic metal nanoparticles onto the surface of a screen-printed carbon electrode, Electrochemistry

Comunications 8, 1375 (2006).

Gold nanoparticle-based redox signal enhancement for sensitive detection of human chorionic gonadotropin hormone Electroanalysis 20,14 (2008)

Gold-Linked Electrochemical Immuno-Assay (GLEIA, PCT/JP2007/56992)

hCG detection by GLEIA

Sample vlume:1 1.5L Lower detection limit: 62.5 pg/mL

Human Chorionic Gonadotropin Hormone standard curves

constructed with buffer () and with serum ()

Potential (V)

0pg/ml

62.5pg/ml

125pg/ml

250pg/ml

500pg/ml

1000pg/m

Typical differential pulse

voltammograms of GLEIA.

Insulin detection by GLEIA

Insulin standard curve constructed with human serum

Sample vlume:1.4L Lower detection limit: 125 pg/mL

HOMA-R fasting blood sugar level (mg/dL) fasting blood insulin level (U/mL)

Application of GLEIA

HOMA-R measurement equipment

HOMA-R is a marker of insulin resistance, metabolic syndrome and diabetics. It is an useful marker to prevent disease progression. We have been developing HOMA-R measurement equipment with the support of Ministry of Economy, Trade and Industry. Blood insulin levels are measured by GLEIA and TAS technologies.

A prototype model of HOMA-R measurement equipment

Measurement chips for insulin and blood sugar

Portable multichannel electrochemical system

Pregnancy, cancer, infectious disease, cardiovascular problems etc.

Clinical Diagnosis

Food safety test

Allergens, food toxins, GMO

Ecological detection

Endocrine disturbing chemicals

Immuno-chromatographic Test Strip and applications

Marketplace in Japan(2006)

$250 million

Influenza diagnosis kits

in Japan

$110 million

Problem Relative low sensitivity around 1 ng/mL Semi-quantitative

Advantage User friendly format, easy to use Rapid to obtain test result Less interface due to chromatographic separation Relative low cost

Suit with screening or POC tests

Applications

Normal strip Enhancer-based strip

Enhancement Assay for Immunochromatographic test strip

PCT/JP2006/323617

Apply sample solution to a

conjugation pad

Antigens react with nanogold-

labeled antibodies and

complex flow to capture antibodies

Conjugation pad Test line

Control line Absorption pad

Capture antibody on test line

Antibody for control line Nanogold labeled antibody

antigen

Antigen-antibody

complex

flow

The complex of antigen and nanogold labeled antibodies accumulated and a higher amount of

gold nanoparticles in close proximity were brought in the enhanced test strip. As a result, higher

Intensity of the localized surface plasmon resonance absorbance of the nanogold particles are

able to obtain and a distinct red color can be easily observed.

Enhancement of sensitivity

Normal and the enhancer-based strips for the detection of hCG in PBS

0

50

1000 100 ontrol

( 1

Normal strip

40

30

20 Inte

nsi

ty

10

10 1 Concentration of hCG (pg/ml)

Enhancer-based strip

In PBS

The lower limit of detection was 100pg/ml

in the normal strip, whereas it was 1pg/ml

in the enhancer-based strip.

pg/ml pg/ml

Normal strip Enhancer-based strip

Test line

Control line

1000 1000 10 10 Control Control

100 1 1 100

In serum

The lower limit of detection was 100pg/ml

in the normal strip, whereas it was 10pg/ml

in the enhancer-based strip.

Production list

Object Species Significance Lower limit of

detection

PSA human Prostate cancer marker 2 ng/mL

hCG human Pregnancy maker 1 pg/mL

Insulin human Metabolic syndrome marker 125 pg/mL

Hemoglobin human Gum disease marker 30 ng/mL

HbA1c human Blood glucose level marker 0.25 ng/mL

Ig E human Maker of allergy to cedar

pollens, mite and cat hair Level 2

sIgA canine Stress marker 20 ng/mL

Cortisol human Stress marker 8 ng/mL

Testosterone human Climacteric marker 125 pg/mL

E. coli E. coli Microbial contamination OD600 0.3

Environmental protection heavy metal

pesticide,herbecide

VOC, odorants

endocrine disruptors

waste water control

pathogenic bacteria

Security explosives

bio/chemical weapons

criminal trace

Food production control taste (tongue, nose)

freshness

key chemicals

(sugar, amino acids etc.)

fermentation control

Food safety pesticide, herbecide

pathogenic bacteria/virus

BSE, chicken flu virus

carcinogens, allergy, GMO

Agriculture/ identification of breed

(rice, wagyu, tuna--- )

seed preservation

(virus free--)

soil and compost test

Bioelectronics/energy biotransistor

biocomputer

biobattery

sustainable system

Drug screening bioassay

DNA/protein chip

alternative to animal tests

Medical diagnosis biochemical tests

cancer

life-style disease

allergy

gene

pathogens

Healthcare mental stress

rapid tests for

blood/urine

point of care

testing

Biosensors application for

medical/environmental/food monitoring

Related Publications 1)Electrochemical DNA quantification based on aggregation induced by Hoechst 33258, M.Kobayashi, T.Kusakawa, M.Saito, S.Kaji, M.Oomura, S.iwabichi, Y.Morita, Q.Hasan and E.Tamiya Electrochemistry Communications 6(4) 337-343(2004) 2)Electrochemical coding of single nucleotide polymorphisms by monobase modified gold nanoparticles, K.Kerman, Y.Morita, Y.Takamura, M.Ozsoz, E.Tamiya, Analytical Chemistry, 76, 1877-1884 (2004) 3)Recent trends in electrochemical DNA biosensor technology(Review), K. Kerman, M. Kobayashi, E.Tamiya, Meas.Sci.Technol. 15, R1-R11 (2004) 4)Electrochemical genosensor for the rapid detection of GMO using loop-mediated isothermal amplification, Minhaz Uddin Ahmed, Masato Saito, M. Mosharraf Hossain, S. Ramachandara Rao, Satoshi Furui, Akihiro Hino, Yuzuru Takamura, Masahiro Takagi and Eiichi Tamiya, Analyst, 134, 966-972 (2009) 5)Nanomaterial-based electrochemical biosensors for medical applications, Kagan Kerman, Masato Saito, Eiichi Tamiya, Shohei Yamamura, Yuzuru Takamura, Trends in Analytical Chemistry, 27(7) 585-592(2008) 6)Electrochemical Biosensors for Medical and Food Applications, Minhaz Uddin Ahmed, Mohammad Mosharraf Hossain, Eiichi Tamiya, Electroanalysis, 20(6), 616-626, 2008 7)Electrochemical DNA biosensor using disposable electrochemical printed (DEP) chip for the detection SNPs from unpurified PCR amplicons, Minhaz Uddin Ahmed, Koutarou Idegami, Miyuki Chikae, Kagn Kerman, Piyasak Chaumpluk, Shouhei Yamamura and Eiichi Tamiya Analyst 132, 431-438 (2007) 8)Electrochemical Genosensor Based on Peptide Nucleic Acid-Mediated PCR and Asymmetric PCR Techniques: Electrostatic Interactions with a Metal Cation, Kerman, K.; Vestergaard, M.; Nagatani, N.; Takamura, Y.; Tamiya, E., Analytical Chemistry, 78(7), 2182-2189 (2006) 9)Novel electrochemical identification and semi quantification of bovine constituents in feedstuffs, P. Chaumpluk et al., Science and Technology of Advanced Materials 7 (2006) 263269 10) Direct fabrication of catalytic metal nanoparticles onto the surface of a screen-printed carbon electrode, Electrochemistry Comunications 8, p1375-1380 (2006). 11) An electrochemical approach for the detection of HIV-1 protease, K. Kerman et al. The Royal Society of Chemistry 2007 Chem. Commun., 2007, 38293831 12) Accumulation of amplified target DNAs using thiol/biotin labeling, S1 nuclease, and ferrocenestreptavidinmagnetic system and a direct detection of specific DNA signals with screen printed gold electrode, P. Chaumpluk et al., Science and Technology of Advanced Materials 8 (2007) 323330 13) Amyloid- detection with saccharide immobilized gold nanoparticle on carbon electrode, M. Chikae et al., Bioelectrochemistry 74 (2008), 116-123 14) Gold nanoparticle-based redox signal enhancement for sensitive detection of human chorionic gonadotropin hormone, Koutarou Idegami et al., Electroanalysis 20, (2008), 14-21 15) Semi-real time electrochemical monitoring for influenza virus RNA by reverse transcription loop-mediated isothermal amplification using a USB powered portable potentiostat,N. Nagatani et al., Analyst, DOI: 10.1039/c1an15638a (2011)