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.

R & D of Biosensors High Sensitive Immunochromatography

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
PCT/JP2006/323617

DEP-Chip series for electrochemical biosensors

Electrochemical DNA sensors (JP3642574) Gold-Linked Electrochemical Immuno Assay

GLEIA, PCT/JP2007/56992)

Disposable Type Screen Printed Electrodes DEP-Chip Series

Square working electrode SP-N, SP-P (carbon) SR-N (gold) Round working electrode EP-N (carbon) EP-PP (carbon, WE with ring and dam) ER-N (gold)

PCR-P01

mass production/disposal/low cost Possible customized fabrication

One drop test

Adapted size for PCR tube

Printable electrodes based biosensors

Gene sensing Pathogenic bacteria/virus Salomonella, Ecoli O-157, Hepatis virus B, Influ virus Food safety GMO, Meat quality SNPs ApoE, ADH
Antibody sensing Gold Linked Electrochemical Immuno Assay : GLEIA hCG hormon : 36 pg/mL (612 fM) insulin : 125pg/mL

Enzyme activity sensing Acetylcholine esterase inhibition Organophosphate pesticide : 0.01 ppm HIV protease inhibition

Original principle of DNA sensor

Japanese patent

(Low concentration of DNA or Before amplification)

(High concentration of DNA or After amplification)

Examples of various DNA targets detected

Salmonella invA gene (PCR)
M
1 0.8 0.6 0.4 0.2 0 100

Genetically-modified corn (LAMP)

M a b c
2.5 2 1.5 1 a b c

a b

negative

negative positive
300 400 500 600 700 800

positive
200 300 400 500 600 700

0.5 0 200

O157 gene (LAMP)

M a b c d e
2.5 2 1.5 c 1 0.5 0 100 d positive e 200 300 400 500 600 700 a

Hepatitis B virus gene from blood sample (Nested PCR)

M a b c
2 1.6 1.2 0.8 0.4 0 200 a b c
b

negative

negative positive
300 400 500 600 700 800

Rapid detection for screening of influenza A virus: microfluidic RT-PCR chip and electrochemical DNA sensor
Analyst,136, 2064 (2011)

Semi-real time electrochemical monitoring for influenza virus RNA by RT-LAMP using USB powered portable potentiostat
Analyst, 2011, Advance ArticleDOI: 10.1039/C1AN15638A BDT DEP chip BDT miniSTAT

Gold-Linked Electrochemical Immuno-Assay

(GLEIA, PCT/JP2007/56992)
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

The voltammetric measurement was performed.

DPV* : differential pulse voltanmetry

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)

hCG detection by GLEIA

Human Chorionic Gonadotropin Hormone standard curves constructed with buffer () and with serum ()
Sample vlume:1 1.5L Lower detection limit: 62.5 pg/mL 1000pg/m

Typical differential pulse voltammograms of GLEIA.

500pg/ml 250pg/ml 125pg/ml 62.5pg/ml 0pg/ml

Potential (V)

Insulin detection by GLEIA

Insulin standard curve constructed with human serum

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

Application of GLEIA HOMA-R measurement equipment

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

Immuno-chromatographic Test Strip and applications

Marketplace in Japan(2006) $250 million Influenza diagnosis kits in Japan $110 million

Advantage
Applications Clinical Diagnosis Pregnancy, cancer, infectious disease, cardiovascular problems etc. Food safety test Allergens, food toxins, GMO Ecological detection Endocrine disturbing chemicals Suit with screening or POC tests
User friendly format, easy to use Rapid to obtain test result Less interface due to chromatographic separation Relative low cost

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

Enhancement Assay for Immunochromatographic test strip

antigen
Nanogold labeled antibody Apply sample solution to a conjugation pad Conjugation pad Antigens react with nanogoldlabeled antibodies and complex flow to capture antibodies

PCT/JP2006/323617
Capture antibody on test line Antibody for control line

Test line

Control line

Absorption pad

Antigen-antibody complex

flow

Normal strip

Enhancer-based strip

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
Normal strip Enhancer-based strip 50 40 Intensity 30 20 10 0 1000 100 1 ontrol 10 Concentration of hCG (pg/ml)
Normal strip Enhancer-based strip

(1

Control line

Test line

1000 100

10 1

Control 1000 100

10 1

Control

In PBS The lower limit of detection was 100pg/ml in the normal strip, whereas it was 1pg/ml in the enhancer-based strip.

In serum The lower limit of detection was 100pg/ml in the normal strip, whereas it was 10pg/ml in the enhancer-based strip.

pg/ml

pg/ml

Production list
Object
PSA

Species
human

Significance
Prostate cancer marker

Lower limit of detection

2 ng/mL

hCG
Insulin Hemoglobin HbA1c Ig E

human
human human human human

Pregnancy maker
Metabolic syndrome marker Gum disease marker Blood glucose level marker Maker of allergy to cedar pollens, mite and cat hair

1 pg/mL
125 pg/mL 30 ng/mL 0.25 ng/mL Level 2

sIgA
Cortisol

canine
human

Stress marker
Stress marker

20 ng/mL
8 ng/mL

Testosterone
E. coli

human
E. coli

Climacteric marker
Microbial contamination

125 pg/mL
OD600 0.3

Biosensors application for medical/environmental/food monitoring

Food production control Medical diagnosis taste (tongue, nose) biochemical tests Healthcare freshness cancer Agriculture/ key chemicals life-style disease mental stress
allergy gene pathogens rapid tests for blood/urine point of care testing

Drug screening

bioassay DNA/protein chip alternative to animal tests

heavy metal pesticide,herbecide VOC, odorants endocrine disruptors waste water control pathogenic bacteria

(sugar, amino acids etc.) identification of breed (rice, wagyu, tuna--- ) fermentation control seed preservation Food safety (virus free--) pesticide, herbecide soil and compost test pathogenic bacteria/virus BSE, chicken flu virus carcinogens, allergy, GMO

Environmental protection Security

explosives bio/chemical weapons criminal trace

Bioelectronics/energy
biotransistor biocomputer biobattery sustainable system

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 ferrocene streptavidinmagnetic 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)