Abstract
Diagnosis and monitoring of complex diseases such as cancer require quantitative detection of multiple proteins. Recent work has shown that when specific biomolecular binding occurs on one surface of a microcantilever beam, intermolecular nanomechanics bend the cantilever, which can be optically detected. Although this label-free technique readily lends itself to formation of microcantilever arrays, what has remained unclear is the technologically critical issue of whether it is sufficiently specific and sensitive to detect disease-related proteins at clinically relevant conditions and concentrations. As an example, we report here that microcantilevers of different geometries have been used to detect two forms of prostate-specific antigen (PSA) over a wide range of concentrations from 0.2 ng/ml to 60 μg/ml in a background of human serum albumin (HSA) and human plasminogen (HP) at 1 mg/ml, making this a clinically relevant diagnostic technique for prostate cancer. Because cantilever motion originates from the free-energy change induced by specific biomolecular binding, this technique may offer a common platform for high-throughput label-free analysis of proteinâprotein binding, DNA hybridization, and DNAâprotein interactions, as well as drug discovery.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Emmert-Buck, M.R. et al. Molecular profiling of clinical tissue specimens: feasibility and applications. Am. J. Pathol. 156, 1109â1115 (2000).
Sander, C. Genomic medicine and the future of health care. Science 287, 1977â1978 (2000).
Whelan, J.P., Kusterbeck, A.W., Wemhoff, G.A., Bredehorst, R. & Ligler, F.S. Continuous flow immunosensor for detection of explosives. Anal. Chem. 65, 3561â3565 (1993).
Devine, P.J. et al. A fiberoptic cocaine biosensor. Anal. Biochem. 227, 216â224 (1995).
Zhu, H. et al. Analysis of yeast protein kinases using protein chips. Nat. Genet. 26, 283â289 (2000).
Davies, H., Lomas, L. & Austen, B. Profiling of amyloid beta peptide variants using SELDI protein chip arrays. Biotechniques 27, 1258â1261 (1999).
Heegaard, N.H. & Kennedy, R.T. Identification, quantitation and characterization of biomolecules by capillary electrophoretic analysis of binding interactions. Electrophoresis 20, 3122â3133 (1999).
Mullett, W., Lai, E.P. & Yeung, J.M. Immunoassay of fumonisins by a surface plasmon resonance biosensor. Anal. Biochem. 258, 161â167 (1998).
Rubio, I., Buckle, P., Trutnau, H. & Wetzker, R. Real-time assay of the interaction of a GST fusion protein with a protein ligate using resonant mirror technique. Biotechniques 22, 269â271 (1997).
Nicholson, S., Gallop, J.L., Law, P., Thomas, H. & George, A.J. Monitoring antibody responses to cancer vaccination with a resonant mirror biosensor. Lancet 353, 808 (1999).
Ostroff, R.M. et al. Fixed polarizer ellipsometry for simple and sensitive detection of thin films generated by specific molecular interactions: applications in immunoassays and DNA sequence detection. Clin. Chem. 44, 2031â2035 (1998).
Piehler, J. et al. Label-free monitoring of DNAâligand interactions. Anal. Biochem. 249, 94â102 (1997).
Jones, V.W., Kenseth, J.R., Porter, M.D., Mosher, C.L. & Henderson, E. Microminiaturized immunoassays using atomic force microscopy and compositionally patterned antigen arrays. Anal. Chem. 70, 1233â1241 (1998).
Blonder, R. et al. Application of redox enzymes for probing the antigenâantibody association at monolayer interfaces: development of amperometric immunosensor electrodes. Anal. Chem. 68, 3151â3157 (1996).
Dahint, R., Bender, F. & Morhard, F. Operation of acoustic plate mode immunosensors in complex biological media. Anal. Chem. 71, 3150â3156 (1999).
Bock, J.L. The new era of automated immunoassay. Am. J. Clin. Pathol. 113, 628â646 (2000).
Fritz, J. et al. Translating biomolecular recognition into nanomechanics. Science 288, 316â318 (2000).
Raiteri, R., Nelles, G., Butt, H.-J., Knoll, W. & Skladal, P. Sensing of biological substances based on the bending of microfabricated cantilevers. Sensors and Actuators B 61, 213â217 (1999).
Wu, G. et al. Origin of nanomechanical cantilever motion generated from biomolecular interactions. Proc. Natl. Acad. Sci. USA 98, 1560â1564 (2001).
Hansen, K.M. et al. Cantilever-based optical deflection assay for discrimination of DNA single nucleotide mismatches. Anal. Chem. 73, 1567â1571 (2001).
Madou, M. Fundamentals of microfabrication. (CRC Press, New York; 1997).
Christensson, A. et al. Serum prostate specific antigen complexed to α1-antichymotrypsin as an indicator of prostate cancer. J. Urol. 150, 100â105 (1993).
Lilja, H. Significance of different molecular forms of serum PSA. Urol. Clin. North Am. 20, 681â686 (1993).
Polascik, T.J., Oesterling, J.E. & Partin, A.W. Prostate specific antigen: a decade of discoveryâwhat we have learned and where we are going. J. Urol. 162, 293â306 (1999).
Moulin, A.M., O'Shea, S.J. & Welland, M.E. Micro-cantilever biosensors. Ultramicroscopy 82, 23â31 (2000).
Lavrik, N.V. et al. Enhanced chemi-mechanical transduction at nanostructured interfaces. Chem. Phys. Lett. 336, 371â376 (2001).
Ji, H.F. et al. A novel self-assembled monolayer (SAM) coated microcantilever for low level caesium detection. Chem. Commun. 6, 457â458 (2000).
Staros, J.V. N-Hydroxysulfosuccinimide active esters: bis(N-hydroxyssuccinimide) esters of two dicarboxylic acids are hydrophilic, membrane impermeant, protein crosslinkers. Biochemistry 21, 3950â3955 (1982).
Knoller, S., Shpungin, S. & Pick, E. The membrane-associated component of the amphiphile-activated, cytosol-dependent superoxide-forming NADPH oxidase of macrophages is identical to cytochrome b559. J. Biol. Chem. 266, 2795â2804 (1991).
Waugh, S.M., DiBella, E.E. & Pilch, P.F. Isolation of a proteolytically derived domain of the insulin receptor containing the major site of crosslinking/binding. Biochemistry 28, 3448â3455 (1989).
Katz, E.Y. A chemically modified electrode capable of a spontaneous immobilization of amino compounds due to its functionalization with succinimidyl groups. J. Electroanal. Chem. 291, 257â260 (1990).
Acknowledgements
This work was supported by the Innovative Molecular Analysis Technologies (IMAT) program of the National Cancer Institute (NIH) (Grant R21 CA86132). G.W. and A.M. would also like to thank the Engineering Program of the DOE Basic Energy Sciences (Grant DE-FG03-98ER14870). K.H., H.J., and T.T. were supported by the Office of Biological and Environmental Research (OBER), US Department of Energy under contract DE-AC05-96OR22464 with Oak Ridge National Laboratory, managed by Lockheed Martin Energy Research Corporation.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wu, G., Datar, R., Hansen, K. et al. Bioassay of prostate-specific antigen (PSA) using microcantilevers. Nat Biotechnol 19, 856â860 (2001). https://doi.org/10.1038/nbt0901-856
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1038/nbt0901-856
This article is cited by
-
Design and evaluation of an implantable MEMS based biosensor for blood analysis and real-time measurement
Microsystem Technologies (2023)
-
Label-free PSA electrochemical determination by seed-mediated electrochemically-deposited gold nanoparticles on an FTO electrode
Journal of Solid State Electrochemistry (2022)
-
Nanomechanical sensor for rapid and ultrasensitive detection of tumor markers in serum using nanobody
Nano Research (2022)
-
Magnetic supercluster particles for highly sensitive magnetic biosensing of proteins
Microchimica Acta (2022)
-
A microwell-based impedance sensor on an insertable microneedle for real-time in vivo cytokine detection
Microsystems & Nanoengineering (2021)