Immunoassays: Our most significant accomplishment is a unique
homogeneous assay for macromolecules with sensitivity sufficient
to use in detection of antibodies, a first in this field. Other
areas under developoment include fluorescence polarization areas.
homogeneous chemilluminescence immunoassays. steric inhibition
immunoassays requiring no immunological conjugate synthesis. new
conjugation approaches for antibody-enzyme complex formation, and
new methods for improving sensitivity of classical techniques.
Protein Interaction at Surfaces: Making use of the binding of
protein to latex particles, a new assay for nonspecific protein
has been developed that is an order of magnitude more sensitive
than competing assays at no cost in increased complexity.
Protein-surface interaction is of great importance to fields as
diverse as blood implant materials. dental plaque formation,
pipeline biological fouling, and ship surface fouling. We are
studying the fundamental effects of the surface parameters and the
protein characteristics on the kinetics, thermodynamics, and
conformation effects of this strong, ubiquitous interaction.
Protein/enzyme Modification: A wide variety of protein
modifications has been carried out over a number of years. These
include: binding of antibodies to a variety of surfaces, to
enzymes, to small molecules including radiosensitizers, thiolating
agents, various coupling agents, avidin, biotin and others; enzyme
modifications, both active site directed and non-specific reagents
for isomerases, aldolases, penicillinases, and fucosidases.
Enzyme Mechanisms: Focusing on the isomerases, aldolases and
fucosidase, classical mechanism probes such as pH profiles,
primary,secondary, and solvent isotope effects, intermediate
trapping experiments, active site mapping, etc. have been used to
elucidate molecular mechanisms of these enzymes.
Anomeric Specificity: The anomeric specificity of a number of
glycolytic enzymes was determined. A novel rationale for the
specificity patterns observed was developed postulating a role in
regulation of glycolysis.
1990-1991 Enzyme Cascade Amplification for Detection of Low
Level Drugs. Ben Franklin Advanced Technology
Center. Total Costs= 67,130
1988-1991 LBI Liaison Program $15,000
1988-1989 Immunoassay Development for Microdiagnostics
Instrumentation. Ben Franklin Advanced Technology
Center. Total costs= $35,400.
1986-1987 Laser Based Fluoroimmunoassay. Ben Franklin
Advanced Technology Center. Total costs=
1985-1986 Laser Excitable Fluorophores. Laser Biotechnology
inc. Total costs= $11,500.
1984-1985 Monoclonal Antibodies for Radiosensitizer
Transport. Ben Franklin Advanced Technology
Center. Co-P.I. with N. D. Heindel. Total
1981-1984 Enzyme Immunoassay Conjugate Synthesis
National Institutes of Health
Direct costs= $170,000, Total costs= $260,000
1979-1980 Synthesis of Enzyme-Antibody Conjugates
Total costs= $10,000
1976-1979 Anomeric Specificity and 1-Epimerase Activities
National Institutes of Health
Direct costs= $78,300 Total costs= $120,000
1973-1975 Anomeric Specificity and 1-Epimerase Activities
National Science Foundation
Total Costs= $44,000
1973-1974 Anomerase Activity of Phosphoglucose Isomerase
Total costs= $6,500
1972-1975 Phosphate Esters of Biological Interest
American Chemical Society PRF Type G
Total Costs= $7500
"An Enzymatic Cycling Procedure for B-NADP+ Generated by
3"-Phosphodiesterase, 2":3"-Cyclic Nucleotide" R. L. Stephon, R.
S. Niedbala, K. J. Schray, and N. D. Heindel, Anal. Biochem. 202,
Book Review Chemical Analysis. V. 117. Application of Fluorescence
in Immunoassay. J. Am Chem Soc. 117, 1009 (1992).
"A Novel Heterobifunctional Linker for Formyl to Thiol Coupling"
N. D. Heindel, H. Zhao, R. A. Egolf, C-H. Chang, K. J. Schray, J.
G. Emrich, J. P. McLaughlin, and D. V. Woo, Bioconjugate Chemistry
2, 427-430 (1991)
"Misanidazole Conjugates of the Colorectal Tumor-Associated
Monoclonal Antibody 17-1A" D. B. Pierce, N. D. Heindel, K.J.
Schray, M. M. Jetter, J. G. Emrich, and D. Woo, Bioconjugate
Chemistry, 1, 314-318 (1990)
"Assays of Avidin(5 ng/ml) and Biotin(100 pg/ml) by Fluorescence
Polarization" K. J. Schray, P. G. Artz, and R. C. Hevey,
Analytical Chem. 60, 853-855 (1988)
"Interactions of Porphyrins and Transfer RNA" N. Foster, A. K.
Singhal, M. W. Smith, N. G. Marcos, and K. J. Schray, Biochim.
Biophys. Acta 950, 118-131 (1988)
"Conformation States of Enzymes Bound to Surfaces" R. K. Sandwick
and K. J. Schray, J. Colloid and Interface Science 121. 1-12
"Separation-Free Dual Solid Phase Enzyme Immunoassay for
Macromolecules" K.J. Schray and R.S. Niedbala Analytical
Chemistry 60. 353-356 (1988)
"The Inactivation of Enzymes Upon Interaction with a Hydrophobic
Latex Surface" R.K. Sandwick and K.J. Schray, J. Colloid and
Interface Science 115,130-138 (1987)
"Radiosensitizer Conjugation to the Carcinoma 19-9 Monoclonal
Antibody" K. P. Borlinghaus, D. A. Fitzpatrick N. D. Heindel, J.
A. Mattis, B. A. Mease, K. J. Schray, D. J. Shealy, H. L. Walton,
Jr., and D. V. Woo, Cancer Research 47, 4071-4075 (1987)
"Macromolecular Attachment as a Metabolic Stabilizer for a Labile
Sensitizer" N. D. Heindel, J. M. A. M. Van Dongen, D. A.
Fitzpatrick, B. A. Mease, and K. J. Schray, J. Pharmaceutical
Sciences 76, 384-386 (1987)
"Further Studies on the Catalytic Mechanism of Human Liver
-L-fucosidase" W. J. White, K. J. Schray, and J. A. Alhadeff
Biochem. Biophys. Acta 912, 132-138 (1987)
"A Controlled Oxidation Synthesis of Substituted Aryl
1-Methyl-4-nitro-5-imidazolyl Sulfones as Candidate
Radiosensitizers" N. D. Heindel, C. J. Lacey, R. Egolf, B. M.
Mease and K. J. Schray, J. Hetero Chem. 23, 1087-1090 (1986)
"Letter to Editor" R. S. Niedbala and K.J. Schray, Clin. Chem.
32, 910 (1986)
"An Enzyme Based Assay for Nanogram Quantities of Biotin and
Avidin" R.S. Niedbala, F. Gergits, III, and K.J. Schray, J.
Biochem. and Biophys. Methods 13. 205-210 (1986)
"Active-site Directed Inactivation of Human Liver -L-Fucosidase
by Conduritol C Trans-epoxide" W. J. White, K. J. Schray, G.
Legler, and J. A. Alhadeff, Biochem. Biophys, Acta 873, 198-203
"Protein Quantitation to 10 ng/ml Concentration by Competitive
Binding to Polystyrene Latices" R.K. Sandwick and K.J. Schray,
Analytical Biochemistry, 147, 210 (1985)
"Estimation of Low Density Lipoprotein by the Friedewald Formula
and by Electrophoresis Compared" R.S. Niedbala, K.J. Schray,
R. Foery, and G. Clement, Clinical Chemistry 31, 1762 (1985)
"Studies of the Catalytic Residues at the Active Site of Human
Liver Fucosidase" W.J. White, K.J. Schray, and J.A. Alhadeff,
Biochem. Biophys. Acta 829, 303 (1985)
"Improved Biotinylation of Glucose-6-phosphate Dehydrogenase
Usding Active Blocking Agents" K.J. Schray, F. Gergits, III, and
R.S. Niedbala, Analytical Biochemistry 149, 225 (1985)
"A Novel Method for Protein Quantitation Using a Competitive
Binding Assay" K.J. Schray, R. Best, A. Decillis, Analytical
Biochemistry 119, 299 (1982)
"Coated Tube Enzyme Immunoassay: Factors Affecting Sensitivity and
Effects of Reversible Protein Binding to Polystyrene" E.E. Howell,
J. Nasser, and K.J. Schray, J. Immunoassay 2, 205 (1981)
"Comparative Inactivation and Inhibition of the Anomerase and
Isomerase Activities of Phosphoglucose Isomerase" E.E. Howell and
K.J. Schray, Molecular and Cellular Biochemistry 37, 101 (1981)
"Inhibition and Inactivation of Liver Aldolase", J.M. Waud, E.
Feldman, and K.J. Schray, Archives of Biochemistry and Biophysics
206, 291 (1981)
"Liver Aldolase Anomeric Specificity" K.J. Schray, E.E. Howell,
J.M. Waud, S.J. Benkovic, and B.A. Cunningham, Biochemistry 19,
"Anomerization of Glucose 6-phosphate: Catalysis by Phosphoglucose
Isomerase", K.J. Schray and E.E. Howell, Archives of Biochemistry
and Biophysics 192, 241 (1979)
"Use of 5-UTP Agarose for Ribonuclease Affinity Chromatography",
G.W. Smith, K.J. Schray, and S.W. Schaffer, Anal. Biochemistry 84,
"Anomerization Rates and Enzyme Specificity for Biologically
Important Sugars and Sugar Phosphates", K.J. Schray and S.J.
Benkovic, Accounts of Chemical Research 11, 136 (1978)
"Anomerization of Glucose 6-Phosphate: pH Dependence and Solvent
Isotope Effects", K.J. Schray and E.E. Howell, Archives of
Biochemistry and Biophysics 189, 102 (1978)
"Phosphomannose Isomerase: Isomerization of the Predicted
Beta-D-Fructose 6-Phosphate", K.J. Schray, J.M. Waud, and
E.E. Howell, Archives of Biochemistry and Biophysics 189, 106
"Phosphoryl Transfer", S.J. Benkovic and K.J. Schray in Transition
States of Biochemical Processes, Eds., R.L. Schowen and R.D.
"The Anomeric Specificity of Glycolytic Enzymes", S.J. Benkovic
and K.J. Schray, Advances in Enzymology 44, 139 (1976)
"Proton Magnetic Relaxation Studies of the Interaction of D-Xylose
and D-Xylitol with D-Xylose Isomerase", J.M. Young, K.J. Schray,
and A.S. Mildvan, J. Biological Chemistry 250, 9021 (1975)
"The Anomeric Form of D-Fructose 1,6-Bisphosphate Used as
Substrate in the Muscle and Yeast Aldolazse Reactions",
K.J. Schray, R. Fishbein, W.P. Bullard, and S.J. Benkovic,
J. Biological Chemistry 250, 4883 (1975)
"The Magnitude of Enzyme Transition State Analog Binding
Constants", K.J. Schyray and J.P. Klinman, Biochem. Biophys.
Research Communications 57, 641 (1974)
"Anomeric Specificity of Phosphofructokinase from Rabbit Muscle"
R. Fishbein, P.A. Benkovic,K.J. Schray, I.J. Siewers, J.J.
Steffens, and S.J. Benkovic, J. Biological Chemistry 249, 6041
"Anomeric Specificity of the Alkaline Form of Fructose
1,6-Diphosphatase from Rabbit Liver", P.A. Benkovic, W.P. Bullard,
M. deMaine, R. Fishbein, K.J. Schray, J.J. Steffens, and S.J.
Benkovic, J. Biological Chemistry 249, 930 (1974)
"Mannose 6-Phosphate: Anomeric Form Used by Phosphomannose
Isomerase and Its 1-Epimerization by Phosphoglucose Isomerase"
I.A. Rose,E.L. O'Connell, and K.J. Schray, J. Biological Chemistry
248, 2232 (1973)
"Catalytic Reactions of Phosphoglucose Isomerase with Cyclic Forms
of Glucose 6-Phosphate and Fructose 6-Phosphate", K.J. Schray,
S.J. Benkovic, P.A. Benkovic, and I.A. Rose, J. Biological
Chemistry 248, 2219 (1973)
"The Chemistry of Phosphoryl Transfer" S.J. Benkovic and K.J.
Schray in The Enzymes, VIII, Ed. P.D. Boyer (1973)
"Inactivaztion of Triose Phosphate Isomerase by D- and L-Glycidol
Phosphate", K.J. Schray, E.L. O'Connell, and I.A. Rose, J.
Biological Chemistry 248,2214 (1973)
"Kinetic and Magnetic Resonance Studies of the Mechanism of
D-Xylose Isomerase: I. Binary and Ternary Complexes with
Manganese(II), Substrates, and Inhibitors", K.J. Schray and A.S.
Mildvan, J. Biological Chemistry 247, 2034 (1972)
"Anomeric Specificity and Mechanism of Two Pentose Isomerases"
K.J. Schray and I.A. Rose, Biochemistry 10, 1058 (1971)
"Mechanisms of Hydrolysis of Phosphate Ester Derivatives of
Phosphoenolpyruvic Acid", K.J. Schray and S.J. Benkovic, J.
American Chemical Society 93,2522 (1971)
"Pentacovalent Phosphorous in the Hydrolysis of
Dibenzylphosphoenolpyruvate" S.J. Benkovic and K.J. Schray, J.
American Chemical Society 91,5653 (1969)
"Metal Ion Catalysis of Phosphoryl Transfer from
Phosphoenolpyruvate" S.J. Benkovic and K.J. Schray, Biochemistry
7, 4097 (1968)
"The Kinetics and Mechanism of Phosphoenolpyruvate Hydrolysis"
S.J. Benkovic and K.J. Schray, Biochemistry 7, 4090 (1968)