James LandersJames P. Landers

Biological, Bioanalytical and Clinical Chemistry

Almost every aspect of the biochemical, biomedical and clinical sciences involves separation of species in complex matrices. Electrophoresis has been a benchmark technique for separation and characterization of biologically-active species. Instead of using conventional slab gel electrophoretic approaches, electrophoresis in micron-scale capillaries using applied fields as high as 30,000 volts, results in unprecedented resolution with unique selectivities and short analysis times. As a result of the microscalar nature of the capillary, only microliters of reagent are consumed by analysis with only a few nanoliters of samples injected for analysis. These characteristics, as well as the ability for on-line detection with laser-induced fluorescence sensitivities in the attomole (10-18 moles) range, made capillary electrophoresis (CE) appealing as a replacement for electrophoretic gels in the biomedical and clinical arenas. We have demonstrated the potential impact of CE on clinical diagnostics through the development of new CE-based assays for measuring kidney function, detecting multiple sclerosis and viral infections, screening for lymphoma, as well as for diagnosing drug abuse and alcoholism.

Figure 1.
A - Schematic of capillary electrophoresis instrumentation. B - CE separation of human serum for the diagnosis of alcoholism.

While the diagnostic impact of standard CE technology is clear, an alternative platform for electrophoresis in microscalar structures has evolved in the form of microchip electrophoresis. The use of microfabricated glass devices containing etched capillary-like channels provides an electrophoretic platform akin to CE but with more flexibility. "Microchip electrophoresis" allows for analysis times to be decreased by an order of magnitude over times achievable by CE (as fast as 10-200 seconds) and two orders of magnitude faster than gel electrophoresis. This provides obvious value to clinical diagnostic laboratories in terms of more rapid turn around time and capability for high throughput screening. We have demonstrated this with the detection T-cell and B-cell lymphoma in a separation remarkably faster than with conventional means.

Figure 2. Demonstration of microchip electrophoresis as a technique for rapid diagnosis of T-Cell lymphoma. Sample T1 shows a negative sample, which is represented by the smear after 100 seconds of separation. T4 is a positive sample with a sharp peak.

With a program focused on the application of miniaturized electrophoretic technology to the clinical and forensic sciences, our current efforts involve broadening the scope of applications for microchip technology. This involves addressing issues associated with integrating functions other than 'separation' onto microchips. For example, we are focused on defining approaches for integrating DNA sample preparation into microchips. PCR amplification of DNA carried out using infrared-mediated thermocycling for rapid on-chip amplification and rapid DNA extraction using microchamber-bound solid phases are two examples of our integration efforts.


Figure 3. A - Demonstration of IR-mediated PCR in a polyimide microchip. Total time necessary for thermocycling was 220 seconds. B - Elution profile of DNA in mSPE chip.

The successful integration of DNA extraction and amplification will lead to the development of an "Integrated Diagnostic" or ID-chip, which we ultimately hope will improve laboratory medicine. Efforts are also underway to 1) define better detection systems using acouto-optic technology, 2) develop multichannel devices for high-throughput analysis using this optical technology, 3) explore proteomic aspects of disease using multi-dimensional microchips for protein separations, and 4) apply the relevant methods to forensic applications.

Representative Publications

Vortex dynamics in confined counter-current shearing flows with applications to mixing. Humphrey, J. A. C., Rosales, J. L., Legendre, L. A., LeDuc, J. P., and Landers, J. P. 2008. Int. J. Heat Fluid Flow, 29(4):1089-1102.

AOTF-based multicolor fluorescence detection for short tandem repeat (STR) analysis in an electrophoretic microdevice. Karlinsey, J. M., Landers, J. P. 2008. Lab Chip 8(8):1285-91.

Towards an integrated microfluidic device for spaceflight clinical diagnostics Microchip-based solid-phase extraction of hydroxyl radical markers. Marchiarullo D. J., Lim J. Y., Vaksman Z., Ferrance J. P., Putcha L., Landers J. P. 2008. J Chromatogr A. 1200(2):198-203.

Protein Determination by Microchip Capillary Electrophoresis Using an Asymmetric Squarylium Dye: Noncovalent Labeling and Nonequilibrium Measurement of Association Constants. Sloat, A. L., Roper, M. G., Lin, X., Ferrance, J. P., Landers, J. P. and Colyer, C. L. 2008. Electrophoresis (in press)

A Rapid Two-Step Method for Enhanced Elution and Preferential Lysis of Cells from Cotton Swabs. Norris, J. V., Cunniffe, H., Ferrance, J. F., and Landers, J. P. 2008. J. Forens. Sci. (in press)

Microchip-based Solid Phase Purification of RNA from Biological Samples. Hagan, K. A., Bienvenue, J. M., Moskaluk, C. A., and Landers, J. P. 2008. Anal. Chem. (in press)

Electrophoretic microfluidic devices for mutation detection in clinical diagnostics. Dewald, A. H., Poe, B. L. and Landers, J. P. 2008. Exp. Opin. Mol. Diag. (in press)

Purification of Nucleic Acids in Microfluidic Devices (review). Wen, J., Legendre, L. A., Bienvenue, J. M., and Landers, J. P. 2008. Anal. Chem. (in press)

Towards a Simplified Microfluidic Device for Ultrafast Genetic Analysis With Sample-In/Answer-Out Capability: Application to T-Cell Lymphoma Diagnosis. Legendre, L. A., Morris, C. J., Bienvenue, J. M., Barron, A. McClure, R., and Landers, J. P. 2008. J. Assoc. Lab. Auto. (in press)

A Low-cost, Low-power Consumption Miniature Laser Induced Fluorescence System for DNA Detection on a Microfluidic Device. Shrinivasan, S., Norris, P. M., Landers, J. P. Ferrance J. P. 2007 Clin. Lab. Med. 27:1 173-181.

A Microchip Sensor for Calcium Determination, Caglar, P., Tunce, S. A., Malcik, N., Landers, J. P. Ferrance, J. P. 2007. Anal. Bioanal. Chem. (in press)

On-chip Pumping for Pressure Mobilization of Focused Zones Following Microchip Isoelectric Focusing. Guillo, C., J. Karlinsey, J. M., Ferrance, J. P., Landers, J. P. 2007. Lab Chip 7:112-118.

Infrared temperature control system for a completely noncontact polymerase chain reaction in microfluidic chips. Roper, M. G., Easley, C. J., and Landers, J. P. 2007. Anal. Chem. Feb 15; 79(4):1294-300.

An active microfluidic system packaging technology. Han, K-H, McConnell, R., Easley, C., Bienvenue, J. M., Ferrance, J. P., Landers, J. P. and Frazier, A. B. 2007. Sens. Act. B. 122: 337-46.

Forensic DNA Analysis on Microfluidic Devices: A Review, Horsman, K. M., Bienvenue, J. M., Blaiser, K. and Landers, J. P. 2007. J. Forens Sci. 52(4):784-99.

Expedited, Chemically-enhanced Sperm Cell Recovery from Cotton Swabs for Rape Kit Analysis. Voorhees, J., Manning, K., Linke, S., Ferrance, J. P., Landers, J. P. 2007. J. Foren. Sci. 52(4):800-5.

Thermal Isolation of Microchip Reaction Chambers for Rapid Non-Contact DNA Amplification. Easley C. J., Humphrey, J. A. C. and Landers, J. P, 2007. J Micromech. Microeng. 17:1758-66.

Microfluidic-based nucleic acid purification in a two-stage, dual-phase microchip containing a reverse phase and a photo-polymerized monolith. Wen, J., Guillo, C., Ferrance, J. P. and Landers, J. P. 2007. Anal. Chem. 79:6135-42.

Microfluidic Chip-based Protein Capture from Human Whole Blood using Octadecyl (C18) Silica Beads for Nucleic Acid Analysis from Large Volume Samples. Wen, J., Guillo, C., Ferrance, J. P. and Landers, J. P. 2007. J. Chrom. A. 1171(1-2):29-36.

A valveless microfluidic sample preparation device for dna extraction and amplification of DNA from nanoliter volume samples using conventional instrumentation. Legendre, L. A., Bienvenue, J. M.. Roper, M. G., Ferrance, J. P., Landers, J. P. 2006. Anal. Chem, 78:1444-1451.

On-chip pressure injection for integration of infrared-mediated DNA amplification with electrophoretic separation. Easley C. J., Karlinsey J. M., Landers J. P. 2006. (Journal Cover) Lab Chip. May;6(5):601-610

DNA extraction using a tetramethyl orthosilicate-grafted photopolymerized monolithic solid phase. Wen, J., Guillo, C., Ferrance, J. P., Landers, J. P. 2006. Anal. Chem, 78:1673-1681.

Unconventional detection methods for microfluidic devices. Viskari P. J., Landers J. P. 2006. Electrophoresis. Apr 27;27(9):1797-1810.

Quenching of Electrochemiluminescence of Tris(2,2’-bipyridine) Ruthenium by Ferrocene and Its Application to Quantitative DNA Detection. Cao, W., Ferrance, J. P., Demas, J. and Landers, J. P. 2006. JACS 14;128(23):7572-8.

Extraction of C-Reactive Protein from Serum On a Microfluidic Chip. Roper, M. G., M. L. Frisk, J. P. Oberlander, J. P. Ferrance, B. J. McGrory, Landers, J. P. 2006. Anal. Chim. Acta, 569: 195–202.

Comparison of DNA Quantitation Assays Using LIF Detection on a Commercial Capillary Electrophoresis Instrument. Guillo, C., Ferrance, J. P., Landers, J. P. 2006. J. Chrom. A 1113(1-2):239-43.

Development of a Human-Specific Real Time PCR Assay for the Simultaneous Quantitation of Total Genomic and Male DNA. Horsman, K. M., Hickey, J. A., Cotton, R. W., Landers, J. P. and Maddox, L. O. 2006. J. Foren. Sci. 51(4):758-65.

Enhanced Elution of Sperm from Cotton Swabs Via Enzymatic Digestion for Rape Kit Analysis. Voorhees, J., Ferrance, J. P., Landers, J. P. 2006. J. Foren. Sci. 51(3):574-79.

A Microchip-based Macroporous Silica Sol-gel Monolith for Efficient Isolation of DNA From Clinical Samples. Wu, Q., Bienvenue, J. M., Giordano, B. C., Hassan, B. J., Kwok, Y. C., Norris, P. M., Landers, J. P. and Ferrance, J. P. 2006., Anal. Chem. 78(16):5704-10.

Multicolor Fluorescence Detection on an Electrophoretic Microdevice using an Acousto-Optic Tunable Filter (AOTF). Karlinsey, J. and Landers, J. P. 2006. Anal. Chem. 78(15):5590-6.

A Low-cost, Low-power Consumption Miniature Laser Induced Fluorescence System for DNA Detection on a Microfluidic Device. Shrinivasan, S., Norris, P. M., Landers, J. P., Ferrance J. P.. 2006 J. Assoc. Lab. Auto. 11(4).

Point-of-care biosensor systems for cancer diagnostics/prognostics. Soper, S. A., Brown, K., Ellington, A., Frazier, B., Garcia-Manero, G., Gau, V., Gutman, S. I., Hayes, D. F., Korte, B., Landers, J. P., Larson, D., Ligler, F., Majumdar, A., Mascini, M., Nolte, D., Rosenzweig, Z., Wang, J., Wilson, D. 2006. Biosens Bioelectron. 5;21(10):1932-42.

Transient Electric Field-driven Sample Zone Heating for Enhanced Fluorescent Labeling of Hydrophilic Proteins Giordano, B. C., Horsman, K., Burgi, D. S., Ferrance, J. P., Landers, J. P. 2006. Electrophoresis 27(7):1355-62.

Chitosan as a Novel Polymer for pH-induced DNA Purification in a Totally Aqueous System: I. High Density Open-Channel Pattern for DNA Purification. Cao, W., Easley, C. J., Ferrance, J. P. and Landers, J. P. 2006. Anal. Chem. 78(20):7222-8.

A Fully-Integrated Microfluidic Genetic Analysis System with Sample in-Answer out Capability. Easley C. J.,, Karlinsey J. M., Bienvenue, J. M., Legendre, L. A., Roper, M. G., Feldman, S. H., Hughes, M. A., Hewlett, E. L., Merkel, T. J., Ferrance, J. P. and Landers, J. P. 2006. Proc Natl Acad Sci U S A. Dec 19;103(51):19272-7. [Editor’s Choice Science Jan. 19, 2007 Vol. 315 Pg 302: Research Highlight Nature Biotech. Vol. 25(1) Jan. 2007; Biosphere in Analytical Chem. Feb. 1, 2007; National Public Radio http://www. earthsky. org/radioshows/51082/new-blood-test-leads-to-early-disease-diagnosis]

Separation of Sperm and Epithelial Cells on Microfabricated Devices: Potential Application to Forensic Analysis of Sexual Assault Evidence. Horsman, K.M., Barker, S.L.R., Ferrance, J.P., Forrest, K.M., Koen, K.A. and Landers J.P. 2005. Anal. Chem. 77(3):742-9. (also see C&E News, Jan.17, 2005, pg 15)

Extrinsic Fabry-Perot Interferometry for Non-Contact Temperature Control of Enzymatic Reactions in Microchips. Easely, Chris, Lindsay A. Legendre, Christopher J. Easley, Thomas A. Wavering, Ferrance, J.P., Landers, J.P. 2005. Anal. Chem. 77(4); 1038-1045.

Evaluation of sieving polymers for fast, reproducible electrophoretic analysis of short tandem repeats (STR) in capillaries. Bienvenue, J.M., Wilson, K., Ferrance, J.P., Landers, J.P. 2005. J. Foren. Sci. 50(4):842-8

Pressure Injection on a Valved Microdevice for Electrophoretic Analysis of Submicroliter Samples. Karlinsey, J., Monahan, J., Marchiarullo, D.J., Ferrance, J.P., Landers, J.P. 2005. Anal. Chem. 77(11):3637-43.

Devices with Thin Membrane Voltage Junctions for Electrospray Mass Spectrometry. Yue, G.E., Jeffery, E.D, Balchunas, C., Easley, C.J., Landers, J.P., Ferrance, J.P., 2005. Lab Chip. 5(6):619-27.

Microchip Laser-Induced Fluorescence Detection of Proteins at Sub-µg/mL Levels Mediated by Dynamic Fluorescent Labeling Under Pseudodenaturing Conditions. Giordano, BC, Jin, LJ, Couch, A.C. and Landers J.P. 2004. Anal. Chem. 76: 4705-4714.

The design and testing of a silica sol–gel-based hybridization array. J.R. Phinney, Conroy, J.F., Hosticka, B., Power, M.E., Ferrance, J.P., Landers J.P., Norris, P.M. 2004. J. Non-Crystalline Solids 350: 39–45.

The performance of a microchip-based fiber optic detection technique for the determination of Ca(II) ions in urine. Caglar, P., Ferrance, J.P. and Landers J.P. 2004. Sensors and Actuators B: Chem. 107(1):24-31.

Fundamentals and practice for ultrasensitive laser-induced fluorescence detection in micro-analytical systems. Johnson, M.J., and Landers J.P. 2004. Electrophoresis 25(21-22):3513-27.

A Microchip-based Proteolytic Digestion System Driven by Electroosmotic Pumping. Jin, LJ, Ferrance, J, Sanders, J, and Landers JP. 2003. Lab. Chip., 3:11 – 18.

Microchip-based purification of DNA from biological samples. Breadmore, M.C., Wolfe, K.A., Arcibal I.G., Leung W.K., Dicks Dana, Giordano, B.C., Power, M.E., Ferrance, J.P., Hosticka, B., Feldman, S., Norris, P.M. and Landers, J.P. 2003. Anal. Chem. 75 (8):1880-1886.

Hydroxypropyl Cellulose as an Adsorptive Coating Sieving Matrix for DNA Separations: Artificial Neural Network Optimization of Polymer and Electrolyte Conditions for Microchip Analysis. Sanders, J.C., Breadmore, M.C., Kwok, Y.C., Horsman, K.M. and Landers, J.P. 2003. Anal. Chem. 75: 986-94.

Capillary electrophoresis with laser-induced fluorescence detection for laboratory diagnosis of galactosemia. Easley CJ, Jin LJ, Presto Elgstoen KB, Jellum E, Landers JP, Ferrance JP. 2003. J. Chromatogr. A. 1004(1-2):29-37.

Developments towards a complete micro-total analysis system for Duchenne muscular dystrophy diagnosis. Ferrance, J., Wu, Q., Giordano, B.C., Hernandez, C., Kwok, Y., Snow, K., Thibodeau, S.N. and Landers J.P. 2003. Anal. Chim. Acta. 500:223-36.