Active bead-linked immunoassay on protein microarrays.

Protein microarrays are becoming a powerful tool in proteome, biochemical, and clinical studies. In addition to the quality of arrayed immobilized probe molecules, sensitivity of the microarray-based assay is highly dependent on the detection technique. Here we suggest four simple techniques for rapid detection of analytes bound to protein microarrays. The techniques employ functionalized magnetic and non-magnetic beads moved to, from, or along the array surface by external forces. In contrast to other labeling techniques actively controlled physical labels: (i) make detection extremely fast to allow microarray reading in seconds; (ii) provide a low background due to active removal of weakly bound beads; and (iii) provide a highly sensitive detection, since one antigen-antibody bond is capable of holding bead immobilized on the array surface. In combination with the electrophoretically assisted active immunoassay we described recently such active reading allows to reduce total indirect immunoassay time to 7-10 min while having sensitivity in the femtomolar concentration range. High speed, sensitivity, and specificity make active bead-linked detection an ideal choice in rapid high-throughput screening and in emergency diagnostics.

[1]  T. Morozova,et al.  Electrophoresis-assisted active immunoassay. , 2003, Analytical chemistry.

[2]  H. Gaub,et al.  A force-based protein biochip , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[3]  A. Lindner,et al.  Quantitative detection of protein arrays. , 2003, Analytical chemistry.

[4]  R. G. Cox,et al.  Slow viscous motion of a sphere parallel to a plane wall , 1967 .

[5]  P. Tabeling,et al.  Aggregation of paramagnetic particles in the presence of a hydrodynamic shear. , 2005, Journal of colloid and interface science.

[6]  Hermann E. Gaub,et al.  Double-chip protein arrays: force-based multiplex sandwich immunoassays with increased specificity , 2004, Analytical and bioanalytical chemistry.

[7]  Sensitive bondforce measurements of ligand-receptor pairs with magnetic beads. , 2005, Biosensors & bioelectronics.

[8]  G. Fonnum,et al.  Characterisation of Dynabeads® by magnetization measurements and Mössbauer spectroscopy , 2005 .

[9]  Marc Fermigier,et al.  Permanently Linked Monodisperse Paramagnetic Chains , 1998 .

[10]  S. Oldenburg,et al.  Base pair mismatch recognition using plasmon resonant particle labels. , 2002, Analytical biochemistry.

[11]  P. Freitas,et al.  High sensitivity detection of molecular recognition using magnetically labelled biomolecules and magnetoresistive sensors. , 2003, Biosensors & bioelectronics.

[12]  K. Ewalt,et al.  Detection of biological toxins on an active electronic microchip. , 2001, Analytical biochemistry.

[13]  C. Kriz,et al.  Magnetic permeability measurements in bioanalysis and biosensors. , 1996, Analytical chemistry.

[14]  R. Merkel,et al.  Energy landscapes of receptor–ligand bonds explored with dynamic force spectroscopy , 1999, Nature.

[15]  Y. Dufrêne,et al.  Implementation of force differentiation in the immunoassay. , 2000, Analytical biochemistry.

[16]  Gil U. Lee,et al.  A biosensor based on magnetoresistance technology. , 1998, Biosensors & bioelectronics.

[17]  V. Morozov,et al.  Electrospray deposition as a method for mass fabrication of mono- and multicomponent microarrays of biological and biologically active substances. , 1999, Analytical chemistry.

[18]  Reinhard Niessner,et al.  Microarrays for the screening of allergen-specific IgE in human serum. , 2003, Analytical chemistry.

[19]  P. Hawkins,et al.  The use of coated paramagnetic particles as a physical label in a magneto-immunoassay. , 2001, Biosensors & bioelectronics.

[20]  R. McDermott,et al.  Ultrasensitive magnetic biosensor for homogeneous immunoassay. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[21]  M. Heller,et al.  Electric field directed nucleic acid hybridization on microchips. , 1997, Nucleic acids research.

[22]  H Schindler,et al.  Cadherin interaction probed by atomic force microscopy. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[23]  R. Huang,et al.  Detection of multiple proteins in an antibody-based protein microarray system. , 2001, Journal of immunological methods.

[24]  M. Heller,et al.  Rapid determination of single base mismatch mutations in DNA hybrids by direct electric field control. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[25]  R. Georgiadis,et al.  Electrostatic surface plasmon resonance: Direct electric field-induced hybridization and denaturation in monolayer nucleic acid films and label-free discrimination of base mismatches , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[26]  V N Morozov,et al.  Electrospray deposition as a method to fabricate functionally active protein films. , 1999, Analytical chemistry.

[27]  H Schindler,et al.  Detection and localization of individual antibody-antigen recognition events by atomic force microscopy. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[28]  V N Morozov,et al.  Immobilization of proteins in immunochemical microarrays fabricated by electrospray deposition. , 2001, Analytical chemistry.

[29]  D. Kriz,et al.  Advancements toward magneto immunoassays. , 1998, Biosensors & bioelectronics.

[30]  Evan Evans,et al.  Chemically distinct transition states govern rapid dissociation of single L-selectin bonds under force , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[31]  S. Loefas,et al.  Immobilization of proteins to a carboxymethyldextran-modified gold surface for biospecific interaction analysis in surface plasmon resonance sensors. , 1991, Analytical biochemistry.

[32]  P Bongrand,et al.  Measuring the Lifetime of Bonds Made between Surface-linked Molecules (*) , 1995, The Journal of Biological Chemistry.

[33]  Mara Prentiss,et al.  Massively parallel adhesion and reactivity measurements using simple and inexpensive magnetic tweezers , 2002 .

[34]  N. V. Avseenko,et al.  Immunoassay with multicomponent protein microarrays fabricated by electrospray deposition. , 2002, Analytical chemistry.

[35]  Paul E. Sheehan,et al.  Design and Performance of GMR Sensors for the Detection of Magnetic Microbeads in Biosensors , 2003 .