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Anup Singh, Ph.D.

 

Sandia National Laboratories

PO Box 969 Livermore, CA 94551-0969

Phone: 925-294-1260, Fax: 925-194-3020

Email: aksingh@sandia.gov

Education:

Ph.D. 1995,
Chemical Engineering, North Carolina State University, Raleigh. Advisors: Prof. Ruben Carbonell and Prof. Peter Kilpatrick

B.Tech. 1990,
Chemical Engineering, Indian Institute of Technology, Bombay, India.

RESEARCH INTERESTS

Microfluidics

My group is involved in developing novel microfluidic systems for analysis of biological molecules. We have adapted a number of biochemical methods to the microfluidic platform for analysis of proteins, peptides and amino acids including SDS-PAGE, native PAGE, isoelectric focusing, chromatography and immunoassays. For analysis of complex samples, we have also developed on-chip sample preparation methods such as dialysis, filtration and preconcentration. For on-chip processing of cells and other biological macromolecules, we are investigating novel dielectrophoretic techniques to manipulate particles. To make these microchips applicable to practical applications, we also specialize in integration of the chips with miniaturized optical, fluidic and electrical components to develop portable devices; an example being a portable prototype for detection of biomarkers of periodontal disease in human saliva. Another application includes studying mechanisms of host-pathogen interaction at single-cell resolution.

Proteomics

We are developing methods to identify proteins and protein complexes in microbial cells such as D. vulgaris. Efforts are currently under way to understand the regulatory networks in the anaerobic bacteria under a variety of stress conditions including heat, pH, and oxygen. To this end a combination of transcriptomics, proteomics and bioinformatics approaches are being applied to identify genes and synthesized proteins and protein complexes involved in stress response pathways.

We are also using two-dimensional gel electrophoresis (2D-GE and DIGE) followed by mass spectrometry to study interaction of bacterial toxins with immune cells. The goal of this effort is to understand the mechanisms of toxicity as well as discover molecular markers that can potentially be used as targets for diagnostics or therapeutics.

Liposomes for Biosensing and Immunoassays

Another area of my research focuses on synthesis of liposomes for a number of applications including immunodiagnostics, biosensing, and analysis of membrane proteins. We have developed glycolipid-bearing liposomes for sensitive detection of bacterial toxins. Proteoliposomes containing an enzyme, horseradish peroxidase and biotin on the surface were developed as generic reagents to amplify signal in biosensing applications.

RESEARCH PROJECTS

Recent Publications

Patents

Invited Lectures

Presentations

In the News

RECENT PUBLICATIONS (1999 - PRESENT)

A.E. Herr, A.V. Hatch, D.J. Throckmorton, H.M. Tran , J.S. Brennan, W.V. Giannobile, A.K. Singh. "Microfluidic Immunoassays as Rapid Saliva-based Clinical Diagnostics." Proc Natl Acad Sci USA, 2007, 104(13), 5268-5273. PDF file

R. Sapra , S.P. Gaucher, J.S. Lachmann, G.M. Buffleben , G. S. Chirica , J.E. Comer, J.W. Peterson, and A.K. Chopra and A.K. Singh, Proteomic analyses of murine macrophages treated with Bacillus anthracis lethal toxin Microbial Pathogenesis, In press.

AV Hatch, AE Herr, DJ Throckmorton, JS Brennan, and AK Singh. "Integrated Preconcentration SDS-PAGE of Proteins in Microchips Using Photopatterned Cross-Linked Polyacrylamide Gels." Analytical Chemistry, 2006, 78(14), 4976 - 4984.

S.R. Chhabra, Q. He, K.H. Huang, S.P. Gaucher, E.J. Alm, Z. He, M.Z. Hadi, T.C. Hazen, J.D. Wall, J. Zhou, A.P. Arkin, A.K. Singh, Global Analysis of Heat Shock Response in Desulfovibrio vulgaris Hildenborough, Journal of Bacteriology, Mar. 2006; Vol. 188, No. 5, p. 1817-1828.

Song, S; Singh, AK On-chip sample preconcentration for integrated microfluidic analysis, Analytical and Bioanalytical Chemistry; January 2006; v.384, no.1, p.41-43.

Barrett, L.M.; Skulan, A.J.; Singh, A.K.; Cummings, E.B.; Fiechtner, G.J. Dielectrophoretic Manipulation of Particles and Cells Using Insulating Ridges in Faceted Prism Microchannels Anal. Chem. 2005, 2005, Nov 1 2005; v.77, no.21, p.6798-6804.

Skulan, A.J.; Barrett, L.M.; Singh, A.K.; Cummings, E.B.; Fiechtner, G.J. Fabrication and Analysis of Spatially Uniform Field Electrokinetic Flow Devices: Theory and Experiment Anal. Chem. 2005, Nov 1 2005; v.77, no.21, p.6790-6797.

Diana Yu, Joanne Volponi, Swapnil Chhabra, C. Jeffrey Brinker, Ashok Mulchandani and Anup K. Singh
Aqueous sol-gel encapsulation of genetically engineered Moraxella spp. cells for the detection of organophosphates,
Biosensors & Bioelectronics, 20(7), 1433-1437, 2005.

Jose M. Moran-Mirabal, Joshua B. Edel, Grant D. Meyer, Daniel J. Throckmorton, Anup K. Singh, and Harold G. Craighead, "Micrometer-Sized Supported Lipid Bilayer Arrays for Bacterial Toxin Binding Studies through Total Internal Reflection Fluorescence Microscopy", Biophysical Journal, July 2005; vol.89, no.1, p.296-305.

AE Herr, DJ Throckmorton, AA Davenport, AK Singh, "On-chip native gel electrophoresis-based immunoassays for tetanus antibody and toxin," Analytical Chemistry, Jan 15 2005; 77(2), 585-590

Amy E. Herr, Anup K. Singh
Photopolymerized Cross-Linked Polyacrylamide Gels for On-Chip Protein Sizing
Analytical Chemistry, 77, 585-590, 2005.

Simon Song, Anup K Singh, Timothy J. Shepodd, Brian J Kirby
Electrophoretic Concentration of Proteins at Laser-Patterned Nanoporous Membranes in Microchips Anaytical Chemistry, 76(15), 4589-4592, 2004.

Jongyoon Han and Anup K Singh,
Miniaturized Isoelectric Focusing and SDS-PAGE Protein Separation
Journal of Chromatography A., 1049 (1-2), 205-209, 2004.

Pathak, S; Singh, AK; McElhanon, JR; Dentinger, PM,
Dendrimer-activated surfaces for high density and high activity protein chip applications, Langmuir, 20(15), 6075-6079, 2004.

Y.J. Song, Y. Yang, C. Medforth, E. Pereira, A.K. Singh, H. Xu, Y. Jiang, C. J. Brinker, F. van Swol, J. A. Shelnutt.
Controlled Synthesis of 2-D and 3-D Platinum Dendrites Using Porphyrin
Photocatalysts, Journal of the American Chemical Society, 126(2), 635-645, 2004.

Simon Song, Anup K Singh, Timothy J. Shepodd, Brian J Kirby
Microchip dialysis of proteins using in situ photopatterned nanoporous polymer
membranes, Analytical Chemistry, 76(8), 2367-2373, 2004.

Eric Cummings and Anup K Singh
Dielectrophoresis in Microchips Containing Arrays of Insulating Posts: Theoretical and Experimental Results
Analytical Chemistry, 75(18), 4724-4731, 2003.

(Ke Wang, Anup K. Singh and John H. van Zanten),
Aggregation Rate Measurements by Zero Angle Time-Resolved Multiangle Laser Light Scattering, Langmuir, 18(6); 2421-2425, 2002.

Daniel J. Throckmorton, Timothy J. Shepodd, Anup K. Singh
Electrochromatography in Microchips: Reversed-phase Separation of Peptides and Amino Acids Using Photo-Patterned Rigid Polymer Monoliths, Analytical Chemistry, 74, 784-789, 2002.

Renèe Shediac, Sarah M. Ngola, Dan J. Throckmorton, Deon S. Anex, Timothy J. Shepodd, Anup K. Singh
Reverse-Phase Electrochromatography of Amino Acids and Peptides Using Porous Polymer Monoliths, Journal of Chromatography A, 925, 251-262, 2001.

(Lital Alfonta, Itamar Willner, Daniel J. Throckmorton and Anup K. Singh),
Electrochemical and Quartz-Crystal-Microbalance Detection of the Cholera Toxin Employing Horseradish Peroxidase- and GM1-functionalized Liposomes, Analytical Chemistry, 73, 5287-5295, 2001.

Anup K. Singh, Eric B. Cummings, Daniel J. Throckmorton
Fluorescent Liposome Flow Markers for Microscale Particle-Image Velocimetry, Analytical Chemistry, 73, 1057-1061, 2001.

Lital Alfonta, Anup K. Singh, Itamar Willner
Liposomes Labeled with Biotin and Horseradish Peroxidase: A Probe for the Enhanced Amplification of Antigen-Antibody or Oligonucleotide-DNA Sensing Processes by the Precipitation of an Insoluble Product on Electrodes, Analytical Chemistry, 73, 91-102, 2001

Anup K. Singh, Suzanne H. Harrison, and Joseph S. Schoeniger
Gangliosides as Receptors for Biological Toxins: Development of Sensitive Fluoroimmunoassays Using Ganglioside-Bearing Liposomes, Analytical Chemistry, 72, 6019-6024, 2000.

R. B. Bhatia, C. J. Brinker, A. K. Gupta, A. K. Singh
An Aqueous Sol-Gel Process for Protein Encapsulation, Chemistry of Materials, 12, 2434-2441, 2000.

Felice C. Lightstone, Maria C. Prieto, Anup K. Singh, Mari Carmen Piqueras, Randy M. Whittal, Mark S. Knapp, Rod Balhorn, and Diana C. Roe
The Identification of Novel Small Molecule Ligands that Bind to Tetanus Toxin, Chemical Research in Toxicology, 13, 356-362, 2000.

A K Singh, A W Flounders, J V Volponi, C S Ashley, K Wally, J S Schoeniger
Development of Sensors for Direct Detection of Organophosphates, Part I: Immobilization, Characterization & Stabilization of Enzymes Acetylcholineesterase & Organophosphate Hydrolase on Silica Supports, Biosensors & Bioelectronics, 14, 703-713, 1999.

A W Flounders, A K Singh, J V Volponi, S C Carichner, K. Wally, A S Simonian, J R Wild, J S Schoeniger
Development of Sensors for Direct Detection of Organophosphates, Part II: Sol-Gel Modified Field Effect Transistor with Immobilized Organophosphate Hydrolase, Biosensors & Bioelectronics, 14, 715-722, 1999.

PATENTS

  1. "Multidimensional Electrophoresis and Methods of Making and Using There of", US Patent Application pending.
  2. "Dialysis on Microchips using thin porous polymer membranes", US Patent Application pending.
  3. "Dielectrophoretic Systems without Embedded Electrodes", U.S Patent Application filed 09/886,165 on June 20, 2001
  4. "Sol-Gel Method For Encapsulating Molecules", U.S Patent 6,495,352 B1.
  5. "Electrokinetic Concentration of Charged Molecules", US Patent 6,428,666 B1.
  6. "Immunodiagnostic Assays Using Liposomes Carrying Labels Thereof On Outer Liposome Surface ", US Patent 5,494,803.

RECENT INVITED LECTURES (2002- Present)

  • New York Academy of Sciences Conference on Oral Based Diagnostics, Oct 2006, Lake Lanier Island, Georgia
  • Gordon Research Conference 2005, Salivary Glands and Exocrine Secretion, Ventura, California February 6-11, 2005
  • Gordon Research Conference on the Physics and Chemistry of Microfluidics, August 21 - 26, 2005, Magdalen College, Oxford, UK
  • Biochemical analysis in microchips using photopatterned porous polymers American Chemical Society (ACS) National Meeting, Anaheim, CA, March 28-April 1, 2004
  • Microchips Containing Engineered Materials for Biochemical Analysis Materials Research Society (MRS) Spring Meeting, San Francisco, April, 2004
  • A novel miniaturized protein concentrator, Annual Electrochemical Society Meeting, San Francisco, Nov 2003.
  • Microfluidic Systems for Proteomics, Nanobiotechnology Center Seminar Series, Cornell University, Ithaca, NY, Sep 9, 2003.
  • Chromatography and Gel Electrphoresis of Proteins and Peptides in Microchips, BioMEMS 2003, San Jose, CA, June 15-17, 2003.
  • Electrochromatography and gel electrophoresis of proteins and peptides in microchips using photopatterned polymer matrices, American Chemical Society Nortwest Regional Meeting, Bozeman, June 2003.
  • Microfluidic Systems for Biochemical Analysis, Analytical Chemistry Invited Seminar, April 17, 2003, UC Riverside.
  • Microfluidic systems for analysis of biological molecules, Bioengineering Seminar Series, MIT, Cambridge, MA, Oct 10, 2002.
  • Electrochromatography in Microchips, Small Talk 2002, San Diego, July 28-31, 2002.

RECENT PRESENTATIONS (2002 - Present)

  • Controlled Synthesis of 2-D and 3-D Platinum Dendrites Using Porphyrin Photocatalysts, Y. Song, Y. Yang, C. Medforth, E. Pereira, A.K. Singh, H. Xu, Y. Jiang, C. J. Brinker, F. van Swol, J. A. Shelnutt, 226th ACS National Meeting, Sept 1-11, NY, NY.
  • Miniaturized Isoelectric Focusing and SDS-PAGE Protein Separation, Jongyoon Han and Anup K. Singh CSBi Computaional and Systems Biology Conference, January 9-10, 2003, MIT, Cambridge, MA.
  • Rapid Deduction of Stress Reponse Pathways in Metal/Radionuclide Reducing Bacteria, Adam Arkin, Alex Beliaev, Inna Dubchak, Matthew Field, Terry Hazen, Jay Keasling, Martin Keller, Vincent Martin, Frank Olken, Anup Singh, David Stahl, Dorothea Thompson, Judy Wall, and Jizhong Zhou
  • A novel miniaturized protein preconcentrator based on electric field-addressable retention and release. Singh, A. K., Throckmorton, D. J., Kirby, B. J., Thompson, A. P., Micro Total Analysis Systems 2002, Nov 10-15, Nara, Japan.
  • Injection of Sample Bands from Open Channels into Packed Separation Columns, Robert H. Nilson, Stewart K. Griffiths and Anup K. Singh, Micro Total Analysis Systems 2002, Nov 10-15, Nara, Japan.
  • Electrokinetic Trapping of Proteins in Cast-To-Shape Matrices, Alexander Artau, Anup, K. Singh, and Timothy J. Shepodd, Micro Total Analysis Systems 2002, Nov 10-15, Nara, Japan.
  • Miniaturization of Protein Separation: Isoelectric Focusing and SDS-PAGE, Jongyoon Han and Anup K. Singh, Micro Total Analysis Systems 2002, Nov 10-15, Nara, Japan.
  • In-situ Fabrication of Dialysis Membranes in Glass Microchannels Using Laser-induced Phase- Separation Polymerization, Brian J. Kirby, Anup K. Singh, Micro Total Analysis Systems 2002, Nov 10- 15, Nara, Japan.
  • Electrochromatography In Chips: Reversed-Phase Separation Of Peptides And Amino Acids Using Polymer Monoliths Photopatterned In Channels Of A Glass Microchip" (Anup K. Singh, Daniel J. Throckmorton, and Timothy J. Shepodd), High Performance Capillary Electrophoresis (HPCE 2002), Stockholm, Sweden, April 13-18, 2002.

IN THE NEWS

Contact Information

Dr. Anup Singh
Biosystems Research Department
Sandia National Laboratories
MS 9292, PO Box 969
7011 East Avenue
Livermore, CA 94551-0969

aksingh@sandia.gov
phone: 925-294-2949
fax: 925-294-3020