CERTIFICATION/REQUIREMENTS

FORM 101Application for a Grant

PART I

Investing in people, discovery and innovationInvestir dans les gens, la découverte et l'innovation

SIGNATURES (Refer to instructions "What do signatures mean?")

TOTAL AMOUNT REQUESTED FROM NSERC

For Strategic Projects, indicate the Target Area and Sub-TargetArea, if applicable.

YuGiven name

Hua-Zhong (Hogan)Initial(s) of all given names

Language of application

Type of grant applied for

2004/07/04

Family name of applicant

Time (in hours per month) to be devoted to the proposed research /activity

Year 1 Year 2 Year 3 Year 4 Year 5

Head of departmentApplicantApplicant's department, university, tel. and fax nos., and e-mail

Form 101 (2003 W)

20

H.Z.Personal identification no. (PIN)

English FrenchX

President of university(or representative)

Dean of faculty

Idea to Innovation

NO If YES to either question a) or b) – Appendices A and B must be completed

Does any phase of the research described in this proposal a) take place outside an office or laboratory, or b) involve an undertaking as describedin Part 1 of Appendix B?

A prototype device incorporating deoxyribosensors for the electrical detection of cancer related plasma proteins

Primary

3504

Secondary

5402

Primary Secondary

1201

Title of proposal

Write a maximum of ten (10) key words that describe this proposal. Use commas to separate them.

Research subject code(s)

1211

Area of application code(s)

Biosensors, Deoxyribosensors, Self-assembled monolayers, Electrochemistry, biochips, Analytical chemistry, Biomedical dignosis, Angiogenin, angiopoietin-2, cytokine

If this proposal involves any of the following, check the box(es) and submit the protocol to the university certification committee.

BiohazardsHumans

It is agreed that the general conditions governing grants as outlined in the NSERC to this application and are hereby accepted by the applicant and the applicant's employing institution.

Program Guide for Professors apply to any grant made pursuant

Chemistry

125,000 0 0 0 0

Date

Research involving : Human pluripotent stem cells Animals

PROTECTED WHEN COMPLETED

The information collected on this form and appendices will be storedin the Personal Information Bank for the appropriate program.

Version française disponible

Simon FraserTel.: (604) 291-5601FAX: (604) 291-3765hzyu@sfu.ca

218450

CO-APPLICANTS' ORGANIZATIONS AND/OR SUPPORTING ORGANIZATIONS (if organization different from page 1)

CO-APPLICANTS

2

I have read the statement "What do signatures on the application mean?" in the accompanying instructions and agree to it.Research/

activity time(hours/month)

Simon Fraser140061, Sen, D 20

Family name and given name of signing officer,title of position, and name of organization Signature

Use additional page(s) if necessary.Form 101 (2003 W), page 2 of 9 Version française disponible

Personal identification no. (PIN) Family name of applicant

PROTECTED WHEN COMPLETED

pursuant to this application and are hereby accepted by the organization.

It is agreed that the general conditions governing grants as outlined in the NSERC Program Guide for Professors, as well as the statements "What do signatures on the application mean?" and "Summary of proposal for public release" in the accompanying instructions, apply to any grant made

PIN, family name and initial(s) Organization Signature

Yu218450

SUMMARY OF PROPOSAL FOR PUBLIC RELEASE (Use plain language.)

Family name of applicantPersonal identification no. (PIN)

218450 Yu

Business telephone no. (optional):

E-mail address (optional):

(604) 291-5601 hzyu@sfu.ca

This plain language summary will be available to the public if your proposal is funded. Although it is not mandatory, you may choose toinclude your business telephone number and/or your e-mail address to facilitate contact with the public and the media about your research.

PROTECTED WHEN COMPLETED Version française disponible

A biosensor is an analytical device incorporating biological materials including enzymes, antibodies, and nucleic acids to detect analytes that have biological importance. The two key processes are the recognition of target molecules by the sensing entities (immobilized molecular probes) and the signal readout (converting binding events into an electrical signal). A recent invention by Sen and co-workers (for example see, J. Am. Chem. Soc. 2002, 124, 4610) shows that electrical conductivity changes resulting from conformational switches within special DNA constructs (deoxyribosensors) induced by molecular analyte binding have the potential for rapid and sensitive electrical sensing of biomedical analytes of interest. Meanwhile, Yu's group is experimenting with different chip-based methodologies for measuring the electrical conductivity changes induced in these sensors upon analyte binding (for example see, Anal. Chem. 2003, 75, 3902). This joint "Idea-to-Innovation (I2I), Phase 1" proposal highlights the application potential of the new generation of DNA-based electrical biosensors, i.e., the development of a practical device that utilizes electroanalytical / electrical diagnostic assays for important biomedical analytes, including cancer-related cytokines in plasma. The major advantages of these deoxyribosensors over conventional sensing materials are their versatility, and characteristically high affinity, specificity, and chemical / structural stability. Specifically, the prototype device proposed here, a microscale electrochemical cell in conjunction with a novel deoxyribosensor chip, will detect and quantitate the cancer-related proteins, human angiogenin (Angn) and angiopoietin-2 (Ang-2), initially within defined buffer solutions and later in biomedical samples. The success of the proposed research will directly lead to the commercialization of a miniaturized, inexpensive, portable electrochemical system for near-patient clinical testing and on-site monitoring.

3

Form 101 (2003 W), page 3 of 9

Second Language Version of Summary (optional).

RESEARCH ACTIVITY SCHEDULE

Personal identification no. (PIN)

218450Family name of applicant

Yu

Description of activitiesAnticipated

starting dateAnticipated

completion date

(Refer to instructions to see if this section applies to your application. Use additional page(s) if necessary.)

Milestone

4 (RPP, SRO)

Design and testing of novel deoxyribosensors for cancer-related proteins, i.e., Angn and Ang-2

2004-10-01 2005-02-28Sensor design and chip fabrication - stage I

Define the limitation of cationic binding method and explore other electrical detection approaches, e.g., covalent and intercalative labeling with redox molecules

2004-10-01 2005-04-30Development of testing methods

Immobilize the newly developed dexyoribonsensors onto gold chips and optimize the surface density/conformation for sensing

2005-03-01 2005-04-30Sensor design and chip fabrication - stage II

Design and construct the electrochemical microcell and test on standarded samples

2005-05-01 2005-07-31Microcell prototyping and chip testing - stage I

Testing of the newly developed dexyribosensor chips and compare with conventional methods

2005-08-01 2005-10-31Microcell prototyping and chip testing - stage II

The final report presenting all the results obtained in this research will be generated by the two teams

2005-11-01 2005-11-30Final report

PROTECTED WHEN COMPLETEDForm 101 (2003 W), page 4 of 9 Version française disponible

125,000 0 012,500 0 0Patenting/consulting fees

0 0 0

0 0 0

0 0 0

1,000 0 0

0 0 0

0 0 0

0 0 0

4,000 0 0

0 0 0

5,000 0 0Senor fabrication/testing

18,000 0 0New sensor design

0 0 00 0 0

0 0 0

12,500 0 0

0 0 0

0 0 0

35,000 0 0

37,000 0 0

PROPOSED PROJECT COSTS

b)

a)

TOTAL PROJECT COSTS

Total support from industry

See instructions for further details.

Year 1 Year 2 Year 3

In-kindCash In-kindCashCash In-kind1) Salaries and benefits

Studentsa)

b) Postdoctoral fellows

c) Technical/professional assistants

2) Equipment or facility

Purchase or rental

Operation and maintenance costs

User fees

d)

d)

Technology transfer activities6)

Field trials

Prototypes

Publication costsa)

b)

5) Dissemination costs

c)

d)

Project-related travelc)

b) Field work

a) Conferences

4) Travel

3) Materials and supplies

a)

b)

c)

a)

b)

c)

Total support from university

Total support from other sources

5 (RPP except RNG, SPG, JIIRP and IRC)Personal identification no. (PIN)

AMOUNT REQUESTED FROM NSERC

Form 101 (2003 W), page 5-RPP of 9 PROTECTED WHEN COMPLETED Version française disponible

Family name of applicant

Yu218450

125,000 0 0

Patenting/consulting fees 0 0

0 0

0 0

0 0

0 0

0 0

0 0

0 0

0 0

0 0

0 0

0 0

0 0

0 0

0 0

Senor fabrication/testing 0 0

0 0

0 0

0 0

New sensor design 0 0

0 0

PROPOSED PROJECT COSTS

b)

a)

TOTAL PROJECT COSTS

Total support from industry

See instructions for further details.

Year 4 Year 5

In-kindCash Cash In-kind1) Salaries and benefits

Studentsa)

b) Postdoctoral fellows

c) Technical/professional assistants

2) Equipment or facility

Purchase or rental

Operation and maintenance costs

User fees

d)

d)

Technology transfer activities6)

Field trials

Prototypes

Publication costsa)

b)

5) Dissemination costs

c)

d)

Project-related travelc)

b) Field work

a) Conferences

4) Travel

3) Materials and supplies

a)

b)

c)

a)

b)

c)

Total support from university

Total support from other sources

5 (RPP except RNG, SPG, JIIRP and IRC)Personal identification no. (PIN)

AMOUNT REQUESTED FROM NSERC

Form 101 (2003 W), page 5-RPP of 9 PROTECTED WHEN COMPLETED Version française disponible

Family name of applicant

Yu218450

0 0

CONTRIBUTIONS FROM SUPPORTING ORGANIZATIONSYear 1 Year 2 Year 3 Year 4 Year 5

Cash contributions to direct costs of research (Transfer amounts to page five (5);except those for the Ship Time program.)

6

218450Personal identification no. (PIN)

Form 101 (2003 W), page 6 of 9 PROTECTED WHEN COMPLETED Version française disponible

Family name of applicant

Yu

section in the NSERC Program Guide for Professors concerning the eligibility of expenditures for the direct costs of research and the regulations

section if you are reporting in-kind contributions or cash contributions for direct and/or indirect costs of research. Submit a separate copy for each supporting organization.Name of supporting organization

Before completing this section, read the instructions for contributions from supporting organizations and consult the Financial Administration

governing the use of grant funds, and the Guidelines for Supporting Organizations concerning the eligibility of in-kind contributions. Complete the

4)

5)

6)

1)

2)

3)

1)

2)

3)

In-kind contributions to direct costs of research

Salaries for scientific and technicalstaff

Donation of equipment, software

Donation of material

Field work logistics

Provision of services

In-kind contributions to indirect costs of research

Use of organization's facilities

Salaries of managerial andadministrative staff

Total of all in-kind contributions

Contribution to university overhead( optional )

Personal identification no. (PIN) Family Name of applicant

218450 Yu

5.1

EXPLANATION AND JUSTIFICATION OF THE PROPOSED EXPENDITURES (1) Salaries and Benefits

a) Students: The amount requested is to support the research of two graduate students. The applied amount,

$18,500 for each graduate student, is based on the current standard in the Department of Chemistry and the Department of Molecular Biology and Biochemistry (MBB) at SFU. Co-supervised by Dr. Yu (Chemistry) and Dr. Sen (MBB), one student will be primarily working on the development of electrochemical testing methods (selected from Department of Chemistry), while the other will focus on the design of new deoxyribosensors (selected from MBB). In the later stage of the project, both of them will be involved in the prototype design and chip testing.

b) Postdoctoral fellow:

The amount requested is to support the research of a postdoctoral fellow (a candidate has been identified and will be hired immediately once the project is funded). The postdoctoral fellow will be responsible for a large part of the DNA/RNA chemistry, as well as the immobilization of deoxyribosensors on chips and structural characterization in order to optimize the sensor performance. In accordance with the NSERC scale, this postdoctoral fellow will be paid $35,000 annually (including 11% benefits). (2) Equipment or facility

a) Purchase or rental The amount requested is to cover the cost of a compact and portable potentiostat and accessories

(e.g., computer and interface) that will be dedicated to the electrochemical measurements described in the project. The model of µAutolabIII from Eco Chemie B.V. will be suitable in terms of the functions (different electrochemical methods), configuration (compact size and friendly user-interface), and the cost (below $10,000). Including the computer and other accessories, the total cost will be about $12,500.

(3) Materials and supplies

1. Sensor design: including relatively expensive, specifically modified, synthetic oligonucleotides ($10,000); and related cost for in vitro selection experiments to design new sensors ($8,000).

2. Chip testing: gold substrates, ultrapure chemicals for modification and electrochemical characterization of sensor electrodes, and related cost to construct/testing the microscale electrochemical cell ($5,000).

(4) Travel

a) Conferences: The amount requested ($4,000) is to assist the research personnel with attendance at biosensor

conferences, as well as other conferences (CSC, ACS) when symposia relevant to our research interests are held. Some additional assistance is expected from the universities through the professional development allowance.

(5) Dissemination costs

Personal identification no. (PIN) Family Name of applicant

218450 Yu

5.2

The amount requested ($1,000) is to assist with the page charges for publications originating from the research after the I.P being protected.

(6) Technology transfer activities

c) Patenting and consulting fees The amount requested for patenting expenses is $10,000, which is less than 50% of the total cost. The differences will be covered by the industrial liaison (LIO) office at Simon Fraser University. We also requested $2,500 as consulting fee to cover the cost of a more detailed market research study for company information and/or business development, as well as the consultation for expertise from Genome BC and other agencies. The industrial liaison office at Simon Fraser University will subsidize the extra cost.

Personal identification no. (PIN) Family Name of applicant

218450 Yu

8

RELATIONSHIP TO OTHER RESEARCH SUPPORT

Yu: The contributions from the university start-up fund and received NSERC equipment grant (2001) made it possible to set up a fully functional laboratory that is essential to carry out the research activities, particularly the preparation of deoxyribosensor chips and electrochemical characterizations, as outlined in this proposal. The awarded NSERC discovery grant (2003-2006) is committed to support the fundamental research of design, preparation, and characterization of DNA-modified surfaces, which focus on the detailed understanding of DNA surface chemistry, i.e., immobilization / hybridization of short oligonucleotides on surfaces from both thermodynamics and kinetics point of view. Based on the recent progress in the construction of a portable and versatile device (based mercury-drop method) for solid-state electrical measurements of semiconducting materials, I also received a Prototype Development Fund from the University / Industry Liaison Office at SFU to support the business development and potential commercialization. An operating grant was also awarded by the World Gold Council to develop DNA microarrays using gold CD-R technology. All these funds have no budgetary overlap with this Idea to Innovation proposal. Sen: The funding requested in this proposal does not overlap with other funding currently available. My NSERC Discovery grant focuses on the generation of new catalytic DNA and RNA molecules, specifically DNA and RNA enzymes that utilize simple cofactors to repair photochemical damage to themselves. That (NSERC) work has essentially no conceptual or budgetary overlap with this application. My CIHR operating grant, due for renewal this fall, was originally funded to explore the creation of deoxyribosensors, to test whether the concept was viable or not. Funding from the CIHR grant was used to generate the first DNA sensors, but the methodology was entirely biochemical, with no effort proposed or made towards chip design or, indeed, the creation of a practical prototype device. This application therefore takes the deoxyribosensor project in a practical, industrial-applicability direction, whereas the CIHR grant focused exclusively on the pure science of DNA and RNA conducting charge and the possibility of the construction of biosensors from DNA and RNA. For my renewal application to CIHR, the proposal will take an entirely new direction, moving substantially away from DNA charge conduction. Both of us (Sen and Yu) are two of the three co-recipients of a two-year, non-renewable NSERC NanoIP grant (the Principal Investigator in this project is Dr. John Bechhoefer, a colleague in the Physics Department at SFU). The work funded by the NanoIP grant has a small and limited overlap with this proposal in terms of the early-stage preparation of standard sensor chips, however, it concerns the use of Conductive Atomic Force Microscopy (CAFM) to study conduction patterns through single adenosine-DNA construct. The goals of the NanoIP funding are pure scientific, non-application oriented studies of the dynamics of individual deoxyribosensor molecules, the nature of ligand binding, and the study of ensembles of deoxyribosensors to see if they display heterogeneous behavior upon binding ligand molecules.

INTELLECTUAL PROPERTY

Family name of applicantPersonal identification no. (PIN)

9 (SRO, CHRP, RNG, CRD, I2I, IRC, CMTC, RPA)

Complete this section if you need to discuss the plans for protecting and disposing of intellectual property arising from the grant. Do not exceed one page.

Version française disponibleForm 101 (2003 W), page 9 of 9 PROTECTED WHEN COMPLETED

218450 Yu

APPENDIX C Referee Suggestions

(Form 101)

Investing in people, discovery and innovationInvestir dans les gens, la découverte et l'innovation

SEND ONEORIGINAL ONLY

DO NOT PHOTOCOPY

Complete Appendix C for all types of grants (except Discovery Grants, Research Tools and Instruments -Category 1 and Major Facilities Access Grants). Read the instructions before completing the appendix.

Date

2004/07/04Family name of applicant Given name Initial(s) of all given names Personal identification no. (PIN)

Yu Hua-Zhong (Hogan) H.Z.Title of proposal

218450

A prototype device incorporating deoxyribosensors for the electrical detection of cancer related plasma proteins

3rd committee reviewer Personal identification no. (PIN)

2nd committee reviewer Personal identification no. (PIN)

NSERC reviewing committee 1st committee reviewer Personal identification no. (PIN)

EArea(s) of expertise

5

DArea(s) of expertise

4

CArea(s) of expertise

3

BArea(s) of expertise

2

AArea(s) of expertise

1Bioelectrochemistry, Physical organic chemistry

Lennox, R.B. (Bruce)ChemistryMcGill University

Montreal, QC CANADA H3A2K6(514) 987-3000bruce_lennox@maclan.mcgill.ca

Molecular analysis of electrochemical processes: electrochemical nanotechnology.

Lipkowski, J. (Jacek)ChemistryUniversity of Guelph

Guelph, ON CANADA N1G 2W1(519) 824-4120 ext 58543 lipkowski@chembio.uoguelph.ca

Materials science and Electrochemistry

Dahn, J.R. (Jeff)PhysicsDalhousie University

Halifax, NS CANADA B4H 4J3(902) 494-2312jeff.dahn@dal.ca

PIN

PIN

PIN

PIN

PIN Lang.

Lang.

Lang.

Lang.

Lang.

Form 101, Appendix C (2003 W) PROTECTED WHEN COMPLETED Version française disponible