Biosensor-based estimation of kinetic and equilibrium constants.

The steady-state affinity constant for the interaction of glutathione S-transferase (GST) with anti-GST immunoglobulin (IgG) was determined by solution-phase equilibrium analysis. A Biacore concentration assay for the determination of free anti-GST IgG was employed giving a Kd value of 6.83 x 10(-10) M. A simple 1:1 solution-phase, equilibrium model approximated the data well. Furthermore, saturation studies showed a maximum occupation of approximately 50%. The choice of affinity-capture ligand, used to anchor anti-GST IgG to the hydrogel, influenced the interaction curves, as evidenced by contact-time-dependent dissociation-phase curves. This was apparent when performing the analysis on anti-mouse Fc-coated surfaces. When the interaction was conducted on a protein A-coated CM5 sensor chip, the interaction conformed well to ideal behavior and was selected for kinetic analysis of the GST interaction.

[1]  L. Christensen.  Theoretical analysis of protein concentration determination using biosensor technology under conditions of partial mass transport limitation. , 1997, Analytical biochemistry.

[2]  D. Diamond,et al.  Development and application of surface plasmon resonance-based biosensors for the detection of cell-ligand interactions. , 2000, Analytical biochemistry.

[3]  D G Myszka,et al.  Kinetic analysis of macromolecular interactions using surface plasmon resonance biosensors. , 1997, Methods in enzymology.

[4]  A. Chaffotte,et al.  Measurements of the true affinity constant in solution of antigen-antibody complexes by enzyme-linked immunosorbent assay. , 1985, Journal of immunological methods.

[5]  R W Glaser,et al.  Antigen-antibody binding and mass transport by convection and diffusion to a surface: a two-dimensional computer model of binding and dissociation kinetics. , 1993, Analytical biochemistry.

[6]  P. Gershon,et al.  Stable chelating linkage for reversible immobilization of oligohistidine tagged proteins in the BIAcore surface plasmon resonance detector. , 1995, Journal of immunological methods.

[7]  J. Quinn,et al.  The use of regenerable, affinity ligand-based surfaces for immunosensor applications. , 1999, Biosensors & bioelectronics.

[8]  L. Nieba,et al.  Competition BIAcore for measuring true affinities: large differences from values determined from binding kinetics. , 1996, Analytical biochemistry.

[9]  A. Minton,et al.  Analysis of mass transport-limited binding kinetics in evanescent wave biosensors. , 1996, Analytical biochemistry.

[10]  G Gauglitz,et al.  Assessment of affinity constants by rapid solid phase detection of equilibrium binding in a flow system. , 1997, Journal of immunological methods.

[11]  P. Stockley,et al.  Quantitation of the Escherichia coli methionine repressor-operator interaction by surface plasmon resonance is not affected by the presence of a dextran matrix. , 1997, Analytical biochemistry.

[12]  D. Winzor,et al.  Interpretation of deviations from pseudo-first-order kinetic behavior in the characterization of ligand binding by biosensor technology. , 1996, Analytical biochemistry.

[13]  Richard O'Kennedy,et al.  Transduction platforms and biointerfacial design of biosensors for 'real-time' biomolecular interaction analysis , 1999 .

[14]  I. Chaiken,et al.  Interpreting complex binding kinetics from optical biosensors: a comparison of analysis by linearization, the integrated rate equation, and numerical integration. , 1995, Analytical biochemistry.

[15]  R. Karlsson,et al.  Experimental design for kinetic analysis of protein-protein interactions with surface plasmon resonance biosensors. , 1997, Journal of immunological methods.

[16]  D G Myszka,et al.  Global analysis of a macromolecular interaction measured on BIAcore. , 1996, Biochemical and biophysical research communications.

[17]  G. Gauglitz,et al.  Affinity characterization of monoclonal and recombinant antibodies for multianalyte detection with an optical transducer , 1997 .

[18]  D. Winzor,et al.  Studies of protein interactions by biosensor technology: an alternative approach to the analysis of sensorgrams deviating from pseudo-first-order kinetic behavior. , 1997, Analytical biochemistry.

[19]  J. Cann,et al.  Demonstration of an upper limit to the range of association rate constants amenable to study by biosensor technology based on surface plasmon resonance. , 1996, Analytical biochemistry.

[20]  P. Edwards,et al.  Determination of association rate constants by an optical biosensor using initial rate analysis. , 1997, Analytical biochemistry.

[21]  R. Karlsson,et al.  Kinetic and Concentration Analysis Using BIA Technology , 1994 .

[22]  P. Schuck,et al.  Kinetics of ligand binding to receptor immobilized in a polymer matrix, as detected with an evanescent wave biosensor. I. A computer simulation of the influence of mass transport. , 1996, Biophysical journal.

[23]  J. Lidholm,et al.  BIACORE analysis of histidine-tagged proteins using a chelating NTA sensor chip. , 1997, Analytical biochemistry.

[24]  J. Schlessinger,et al.  Measurement of the binding of tyrosyl phosphopeptides to SH2 domains: a reappraisal. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Bo Johnsson,et al.  A novel hydrogel matrix on gold surfaces in surface plasmon resonance sensors for fast and efficient covalent immobilization of ligands , 1990 .