Literature References

Order of citations is chronological

• Polarization de la lumiere de fluorescence: Vie moyenne de molecules dans l'etat excite.  Perrin, F.
  J. Phys. Radium 7:390 (1926).
• Fluorescence polarization in immunochemistry.  Dandliker, W.B, DeSaussure, V.A.
  Immunochemistry 7:799-828 (1970).
• Corrected Equations for the Calculation of Protein-ligand Binding Results from Fluorescence Polarization Data.  K.M. Rajkowski and N. Cittanova.
  J. theor. Biol. 93:691-696 (1981).
• Preparation and Application of a Fluorescein-Labeled Peptide for Determining the Affinity Constant of a Monoclonal Antibody-Hapten Complex by Fluorescence Polarization.  W. Jiskoot, P. Hoogerhout, E. Coen Beuvery, J.N. Herron and D.J.A. Crommelin.
  Anal. Biochem.  196, 421-426 (1991).
• Use of synthetic Peptides as Tracer Antigens in Fluorescence Polarization Immunoassays of High Molecular Weight Analytes.  Wei, Ai-Ping, and James N. Herron.
  Anal. Chem.  65, 3372-3377 (1993).
  This University of Utah group used tetramethylrhodamine to label peptides and built an ultrasensitive competitive binding assay.
• Characterization of a Fluorescent Substance P Analog.  M. R. Tota et al.
  Biochemistry 33, 13079-13086 (1994).
  This group at Merck (Rahway, NJ) presents a detailed procedure for labeling a peptide with fluorescein, purifying a cloned receptor, evaluating assay components and building a sensitive fluorescence polarization assay.
• Interaction of [fluorescein-Trp25]Glucagon with the Human Glucagon Receptor Expressed in Drosophila Schneider 2 Cells.  M.R. Tota et al.
  J. Biological Chemistry  26466-26471 (1995).
  The group at Merck worked with fluorescein-labeled glucagon and a cloned human glucagon receptor in membrane preps.
• Fluorescence Polarization Immunoassay of a High-Molecular-Weight Antigen Based on a Long-Lifetime Ru-Ligand Complex.  Terpetschnig, E., H. Szmacinski and J.R. Lakowicz.
  Anal. Biochem.  227:140-147 (1995).
• Metal-Ligand Complexes as a New Class of long-lived Fluorophores for Protein Hydrodynamics. Terpetschnig, E., H. Szmacinski, H. Malak and J.R. Lakowicz.
  Biophysical J. 68:342-350 (1995).
• Fluorescence Anisotropy Applied to Biomolecular Interactions.  D.M.  Jameson and W.H. Sawyer.
  Methods in Enzymology, 246, 283-300 (1995).
  This is a review article covering both theory and applications of fluorescence polarization.
• Components of a Stat Recognition Code: Evidence for Two Layers of Molecular Selectivity.  U. Schindler, P. Wu, M. Rothe, M. Brasseur and S.L. McKnight.
  Immunity 2, 689-697 (1995).
  Tularik authors use fluorescence polarization to conduct saturation binding assays for protein-phosphopeptide interactions.  They constructed competition binding assays, showed specificity using peptides from interferon gamma and IL-4 receptor.  Tracer peptides were labeled at the N-terminal amine by NHS esters of fluorescein.
• Predicting ligand binding to proteins by affinity fingerprinting.  Kauvar, L. M., Higgins, D. L., Villar, H. O., Sportsman, J. R., Engqvist-Goldstein, A., Bukar, R., Bauer, K. E., Dilley, H., and Rocke, D. M.
  Chem. & Biol. 2:107-118.
• Fluorescence Polarization Immunoassay of a High-Molecular-Weight Antigen Using a Long Wavelength-Absorbing and Laser Diode-Excitable Metal-Ligand Complex.  Terpetschnig, E., H. Szmacinski and J.R. Lakowicz.
  Anal. Biochem.  240:54-59 (1996).
• Synthesis and evaluation of Ru-complexes as anisotropy probes for protein hydrodynamics and immunoassays of high-molecular-weight antigens.  Szmacinski, H., E. Terpetschnig and J.R. Lakowicz.
  Biophysical Chemistry 62:109-120 (1996).
• Fluorescence Polarization Assays for the Detection of Proteases and Their Inhibitors.  M.E. Jolley,
  J. Biomolec. Screening.  1, 33-38 (1996).
  Fluorescence polarization expert uses BODIPY dyes to create generic protease assays by labeling casein.  This describes a general assay strategy for all hydrolytic enzymes.  Jolley cites earlier work done at several Japanese pharmaceutical companies with this technology.

• Application of Fluorescence Polarization Assays in High-Throughput Screening.  J.C. Owicki, M. Biros and W. Burton.
  Genetic Engineering News, volume 17, number 19 (1997).
  An overview of the application of HEFP to High Throughput Screening by LJL scientists.
• Specificity of Aminoglycoside Binding to RNA Constructs Derived from the 16S rRNA Decoding Region and the HIV-RRE Activator Region.  Y. Wang, K. Hamasaki and R.R. Rando.
  Biochemistry 36, 768-779 (1997).
  Harvard University group studied interaction aminoglycoside antibiotics to ribosomal RNA and to HIV RNA sequences using fluorescence polarization.  The dissociation constant for tracer binding to an HIV sequence was shown to be 7.6 nM. The label was 5-carboxytetramethylrhodamine.
• Fluorescence Polarization.  J.R. Sportsman, S.K. Lee, H. Dilley and R. Bukar.  "High Throughput Screening" J.P. Devlin, editor.  Marcel Dekker, Inc. New York (1997).
  This review by the South San Francisco biotech company Terrapin (now named Telik) describes their success with fluorescence polarization and discusses color quenching, sensitivity and reagents.
• Measurement of Specific Protease Activity Utilizing Fluorescent Polarization.  L.M. Levine, M.L. Michener, M.V. Toth and C.C. Holwerda.
  Anal. Biochem.  247, 83-88 (1997).
  Monsanto and Searle scientists report novel protease format using peptides with protease-specific scissile bond and two labels (fluorescein and biotin).  Addition of avidin provides the size change to create changes in net polarization.
• A Fluorescent Polarization Based Src-SH2 Binding Assay.  B.A. Lynch, K.A. Loiacono, C.L. Tiong, S.E. Adams and I.A. MacNeil.
  Anal. Biochem. 247, 77-82 (1997).
  This group from ARIAD Pharmaceuticals (Cambridge, MA) reports an FP binding assay for the recognition of the SH2 (Src homology 2) region of phosphotyrosine-containing proteins.  Their tracer was a carboxyfluorescein-labeled pentapeptide containing phosphotyrosine.
• A homogeneous, fluorescence polarization assay for Src-family tyrosine kinases.  Seethala, R. and R. Menzel.
  Anal. Biochem.  253:210-218 (1997).
  This report describes the optimization of various assay parameters and presents the IC50's for 7 different peptides
• A high-throughput STAT binding assay using fluorescence polarization.  P. Wu, M. Brasseur, U. Schindler.
  Anal. Biochem. 249:29-36 (1997).
• Homogeneous GPCR-Ligand Binding Assays in 1536 well Microtiter plates.  Peter Banks, Meng Zhang, Neil Bristol,  poster presentation at the Society for Biomolecular Screening, Baltimore, MD September, 1998.
  The authors, from Advanced Bioconcept Co. and LJL BioSystems, Inc. reported miniaturized GPCR FP assays (5 µl/well) using MC4 and MC5 receptors. The FP assays were conducted using crude membrane preps and employed LJL Acquest.
• A Fluorescence Polarization Competition Immunoassay for Tyrosine Kinases.  Ramakrishna Seethala and R. Menzel.
  Anal. Biochem. 255, 257-262 (1998).
  Bristol-Myers Squibb scientists describe a competitive fluorescence polarization immunoassay for tyrosine kinase product.
• Long-lifetime Ru(II) complexes for the measurement of high molecular weight protein hydrodynamics.  Szmacinski, H., F.N. Castellano, E. Terpetschnig, J.D. Dattelbaum, Lakowicz, J.R. and G. J. Meyer.
  BBA 1383:151-159 (1998).
• Development of Fluorescence Polarization and FRET Assays for Tyrosine and Serine/Threonine Kinases.  Jinzi J. Wu.  Presented at the IBC
  2nd Annual International Conference on Miniaturization Technologies, Berkeley, California, March 4-5, 1999.
  Novartis (Summit, NJ) scientist uses Analyst to develop HEFP assays for kinases.
• BOCILLIN FL, a Sensitive and Commercially Available Reagent for Detection of Penicillin-Binding Proteins.  G. Zhao, T.I. Meier, S.D. Kahl, K.R. Gee and L.C. Blaszczak.
  Antimicrobial Agents and Chemotherapy, 43, 1124-1128 (1999).
  Eli Lilly scientists used Analyst to develop HEFP assay for antibiotic binding proteins.
• Fluorescence Polarization in Homogeneous Nucleic Acid Analysis.  Chen, X., L. Levine and P.-Y. Kwok.
  Genome Research, 9, 492-498 (1999).
  Washington University (St. Louis, MO) scientists use LJL Analyst and HEFP to build an assay to detect point mutations in human DNA.  Single nucleotide polymorphisms are detected by a terminal extension of a primer adjacent to the site of polymorphism, using PCR, primer extension and HEFP.  The system uses as many as 4 different fluorescent dyes in the same well.
• A robust, versatile tyrosine kinase assay for HTS in drug discovery.  Deshpande, S., Mineyev, I., Owicki, J.
  Prog. Biomed. Optics (SPIE)  3603:251-261 (1999).
  Principles of Fluorescence Spectroscopy.  J.R. Lakowicz.  Second edition, 1999.  Kluwer Academic/Plenum Publishers, New York.
  The definitive text on this subject, updated.

• Analysis of Protein-Peptide Interactions by a Miniaturized Fluorescence Polarization Assay Using Cyclin Dependent Kinase 2/Cyclin E as a Model System.  Ilona Kariv, podium presentation at the Society for Biomolecular Screening, Edinburgh, UK September 1999.
  DuPont Scientist uses HEFP and LJL Acquest to build very low volume assay in 1536-well plates.
• 1536-well Screening: The Leap From Proof of Concept to HTS.  Alan Binnie, podium presentation at the Society for Biomolecular Screening, Edinburgh, UK September 1999.
  Selectide group (a division of Hoechst-Marion-Roussel) is among the leaders in HTS minaturization and reported success in developing assays and running screens with LJL Acquest.
• Homogeneous G-protein Coupled Receptor-Ligand Binding Assays for High Throughput and Ultra-high Throughput Screening.  Peter Banks, M. Gosselin and L. Prystay, poster presentation at the Society for Biomolecular Screening, Edinburgh, UK September 1999.
  The authors, from Advanced Bioconcept Co. (Montreal, Canada), reported on fluorescent analogs of deltorphin, dynorphin, NDP-MSH and neurotensin, with corresponding receptors.  The FP assays were conducted using crude membrane preps and employed LJL Analyst.
• Discovery of an Anti-Phosphoserine Antibody for the Development of Fluorescence Polarization Assays of Serine/Threonine Kinases.  Jinzi J. Wu, Q. Pham, D.R. Yarwood, J. Tivade, J. Bernstein, T. Kowski, A. Clark and M.A. Sills, poster presentation at the Society for Biomolecular Screening, Edinburgh, UK September 1999.
  This poster was awarded by the conference as among the top 5% at SBS.  The Novartis (Summit, NJ) scientists describe an antibody with a Kd of 250 pM as determined in the FP assay.  The antibody was used to develop an assay for multiple serine/threonine kinases 384 well plates and for several such enzymes in 1536 plates.
• Simultaneous Assay of Src SH3 and SH2 Domain Binding Using Different Wavelength Fluorescence Polarization Probes.  B.A. Lynch, C. Minor, K.A. Loiacono, M.R. van Schravendijk, M.K. Ram, R. Sundaramoorthi, S.E. Adams, T. Phillips, D. Holt, R.J. Rickles and I.A MacNeil.
  Analytical Biochemistry 275, 62-73 (1999).
  The scientists from ARIAD Pharmaceuticals, Inc. (Cambridge, Massachusetts)
• Homogeneous fluorescence readouts for miniaturized high-throughput screening: theory and practice.  Pope, A.J., U.M. Haupts and K.J. Moore.
  Drug Discovery Today 4:350-362 (1999).
• An Ultra-High Throughput Screening Approach for an Adenine Transferase Using Fluorescence Polarization.  Li, Z., S. Mehdi, I. Patel, J. Kawooya, M. Judkins, W. Zhang, K. Diener, A. Lozada and D. Dunnington.
  J. Biomolecular Screening 5:31-37 (2000).
  The authors used Acquest in 1536 mode with a 6.5 µl final volume, a tracer custom synthesized by LJL (contains Texas Red) and ran it on a 250,000 compound library.  They concluded FP "ideal for ultra-HTS" and preferable to SPA and to a "filtration method" and noted that FP "provided a 10-fold savings in enzyme, thereby reducing the time required to generate essential biological reagents".
• Identification of a High-Affinity Anti-Phosphoserine Antibody for the Development of a Homogeneous Fluorescence Polarization Assay of Protein Kinase C.  Wu, J.J., D.R. Yarwood, Q. Pham, and M.A. Sills.
  J. Biomolecular Screening.  5:23-30 (2000).
  Wu and colleagues conclude their "FP assay is more sensitive and robust than the scintillation proximity assay for PKC."
• Detection of Phosphopeptides by Fluorescence Polarization in the Presence of Cationic Polyamino Acids: Application to Kinase Assays.  Coffin, J., M. Latev, X. Bi and T.T. Nikiforov.
  Anal. Biochem. 278:206-212 (2000).
• High Throughput Fluorescence Polarization: A Homogeneous Alternative to Radioligand Binding for Cell Surface Receptors.  Allen, M., J. Reeves and G. Mellor.
  J. Biomolecular Screening 5:63-69 (2000).
• Development of High Throughput Screening Assays Using Fluorescence Polarization: Nuclear Receptor-Ligand-Binding and Kinase*/Phosphatase Assays.  Parker, G.J., T.L. Law, F.J. Lenoch, and R.E. Bolger.
  J. Biomolecular Screening 5:77-88 (2000).
• Cell Permeable Bcl-2 Binding Peptides: A Chemical Approach to Apoptosis Induction in Tumor Cells.  Wang, J.-L., Z.-J. Zhang, S. Choksi, S. Shan, Z. Lu, C.M. Croce, E.S. Alnemri, R. Korngold and Z. Huang.
  Cancer Research 60:1498-1502 (2000).
  The authors constructed a competitive binding assay for assessing the binding of peptides to the Bcl-2 protein, using a 5 carboxyfluorescein-peptide.  The system was used to evaluate lipid-modified peptides.  The estimated dissociation constant of the tracer peptide for the binding protein was about 340 nM.
• Homogeneous high-throughput screening of G protein-coupled receptors using fluorescence polarization.  Banks, P. and M. Gosselin.
  American Biotechnology Laboratory, pages 28-30, April 2000.
  The authors use FP to demonstrate the utility of their product Fluo-peptide.  A 40 µl competitive binding assay using 1 nM of a fluorescent analog of NDP-alpha MSH and 0.6 to 1.0 nM MC4 receptors was shown to give a delta mP of about 70 mP with good precision.  Displacement curves with three peptide agonists showed a rank order similar to that achieved with a radioisotope filtration assay.  The authors used an LJL Analyst.
• Development of a Homogeneous High-Throughput Fluorescence Polarization Assay for G Protein-coupled Receptor Binding.  P.H. Lee and D.J. Bevis.
  J. Biomolecular Screening 6:415-419 (2000).

• Putting the pieces together:  Contribution of fluorescence polarization assays to small molecule lead optimization.  Keating, S., Marsters, J., Beresini, M., Ladner, C., Zioncheck, K., Clark, K., Arellano, F., and Bodary, S.
  Proc. SPIE 3913:128-137 (2000).
• Fluorescence polarization assays for high throughput screening of G protein-coupled receptors.  Banks, P., Gosselin, M., and Prystay, L.
  J. Biomol. Screening 5:159-168 (2000).
• Fluorescence polarization for monitoring ribozyme reactions in real time.  Singh, K.K., Ruecker, T., Hanne, A., Parwaresch, R. and Krupp, G.
  BioTechniques 29:344-351 (2000).
• Fluorescence polarization and anisotropy in high throughput screening: perspectives and primer. Owicki, J.C.
  J. Biomol. Screening 5:297-306 (2000).
• Impact of a Red-Shifted Dye Label for High Throughput Fluorescence Polarization Assays of G Protein-Coupled Receptors.  Banks, P., Gosselin, M., and Prystay, L.
  Journal of Biomolecular Screening 5:329-334 (2000).
• Real Experiences of uHTS: A Prototypic 1536-Well Fluorescence Anisotropy-Based uHTS Screen and Application of Well-Level Quality Control Procedures.  Turconi, S., Shea, K., Ashman, S., Fantom, K., Earnshaw, D.L., Bingham, R.P., Ulrich M. Haupts, U.M., Brown, M.J.B., and Pope, A.J.
  Journal of Biomolecular Screening 6:275-290 (2001).
• Single Molecule Detection Technologies in Miniaturized High Throughput Screening: Binding Assays for G Protein-Coupled Receptors Using Fluorescence Intensity Distribution Analysis and Fluorescence Anisotropy.  Rüdiger, M., Haupts, U., Moore, K.J., and Pope, A.J.
  Journal of Biomol. Screening 6:29-37 (2001).
• Determination of Equilibrium Dissociation Constants in Fluorescence Polarization.  Prystay, L., Gosselin, M., Banks, P.
  Journal of Biomol. Screening 6:141-150 (2001).
• Use of Fluorescence Polarization Detection for the Measurement of Fluopeptide^™ Binding to G Protein-Coupled Receptors  Gagne, A., Banks, P., and Hurt, S.D.
  Journal of Receptors and Signal Transduction 22:333-343 (2002).
• A Homogeneous Fluorescence Polarization Assay Adaptable for a Range of Protein Serine/Threonine and Tyrosine Kinases.  Gaudet, E.A., Huang, K., Zhang, Y., Huang, W., Mark, D., and Sportsman, J.R.
  Journal of Biomolecular Screening 164-175 (2003).
• A Carboxy-terminal Mutation of the Epidermal Growth Factor Receptor Alters Tyrosine Kinase Activity and Substrate Specificity as Measured by a Fluorescence Polarization Assay.  Beebe, J.A, Wiepz, G.J., Guardarrama, A.G., Berties, P.J. and Burke, T.J.
  Journal of Biological Chemistry 26810-26813 (2003).
• Utilization of Polarization and Time-resolved Fluorescence Resonance Energy Transfer Assay Formats for SAR Studies: Src Kinase as a Model System.  Newman, M., and Josiah, S.
  J. Biomol Screening 9:525-532 (2004).