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Group Research Interests - an overview. James F. Rusling Professor of Chemistry and Pharmacology University of Connecticut and University Health Center. Example 1. Toxicity Screening. Lipophilic Molecule. Cyt P450, O 2. Enzyme-activated molecule. + DNA. Damaged DNA.

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slide1

Group Research Interests - an overview

James F. Rusling

Professor of Chemistry and Pharmacology

University of Connecticut and University Health Center

slide2

Example 1. Toxicity Screening

Lipophilic Molecule

Cyt P450, O2

Enzyme-activated molecule

+DNA

Damaged

DNA

Detect by voltammetry, LC-MS/MS

slide5

Collaboration with Prof. John Schenkman,

Pharmacology, Uconn Health Center

Funding from NIH, NIEHS

slide6

Screening Chemical Toxicity

Enzyme reaction - Incubate:

Reactant + H2O2-->metabolite

Analysis by catalytic SWV or

electrochemiluminescence

RuL2+ = RuL3+ + e-

RuL3+ + DNA-G --> RuL2+ + DNA-G•

slide7

Enzyme/DNA films

Peak increase

measures damage

of DNA by enzyme-

generated

metabolite

slide9

Comparison of toxicity sensors with LC-MS

For DNA damage by methylmethane sulfonate

slide12

Arrays detect in-vitro DNA damage from metabolites of

different enzymes with DNA/enzyme films

Rel. turnover rate,

1/min (nmol Enzyme)

Drug development

applications

slide13

Sensors for oxidative stress via oxidized DNA

Os-PVP

PSS

Pyrolytic Graphite

Ru-PVP

ECL detection in films:

Lynn Dennany, Robert J. Forster, Blanaid White, Malcolm Smyth and James F. Rusling, J. Am. Chem. Soc., 2004, 126, 8835-8841.

slide14

Single-Walled Carbon Nanotube

Forests: Antigen-Antibody Sensing

SPAN or

Nafion

Chattopadhyay, Galeska, Papadimitrakopoulos, J. Am. Chem. Soc. 2001, 123, 9451.

End COOH groups allow chemical attachment to proteins (antibodies)

slide15

(b)

(a)

AFM of SWNT forest with and without antibody attached

Collaboration with F. Papadimitrakopoulos (ARO funded)

SWNT + antibody

(EDC coupling)

SWNT

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H2O2

HRP

HRP

HRP

HRP

HRP

HRP

Ag

Ag

Ab1

Ab1

Sandwich Assay for Human Serum Albumin

Ab2

HPR

SWNT forest

Apply E measure I

slide17

Detection of Human Serum albumin in

10 mL drops on SWNT forest immunosensor

LOD ~ 10 pmol/mL

slide18

Styrene Epoxidation Catalyzed by

Myoglobin-Polyion Films

in Microemulsions (NSF funded)

Stable in microemulsions

slide20

Additional Current Projects

• “Green” chiral synthesis using enzyme films in microemulsions, nanoparticle catalysts (NSF)

• Redox chemistry of complex photosynthetic proteins

in membrane films (with H. Frank, USDA funded)

• detection of Lyme disease vectors (proteins) in tick saliva (with Uconn Health Center)

• enzyme activity for generating toxic metabolites (with Uconn Health Center)

• EC-Laser fluorescent detection of DNA damage

• arrays for anticancer biomarkers (NIH)

• molecular dynamics of film construction

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