Extracting kinetic rate constants from surface plasmon resonance array systems

Rich, Rebecca L.; Cannon, Michelle J.; Jenkins, Jerry; Pandian, P. Saravana; Sundaram, S.; Magyar, Rachelle A.; Brockman, Jennifer M.; Lambert, Jérémy; Myszka, David G.

doi:10.1016/j.ab.2007.08.017

Cited by 70 publications

(61 citation statements)

References 10 publications

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“…Indeed, one would have expected that multiple interactions between one mAb and two antigens might occur simultaneously. Similar cases of good adequation between global fits with a simple model and experimental data that were expected to deviate from such a simple model have already been reported (Svitel et al, 2003;Rich et al, 2008). In our case, it is more likely that additional binding events leading to the formation of a 2:1 stoichiometry complex may occur on a different time-scale than the one explored in both non-optimized and optimized experiments.…”

Section: Preliminary Experimental Data and Optimizationsupporting

confidence: 75%

Online optimization of surface plasmon resonance‐based biosensor experiments for improved throughput and confidence

Crescenzo

Woodward

Srinivasan

2008

J of Molecular Recognition

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The emergence of surface plasmon resonance-based optical biosensors has facilitated the identification of kinetic parameters for various macromolecular interactions. Normally, these parameters are determined from experiments with arbitrarily chosen periods of macromolecule and buffer injections, and varying macromolecule concentrations. Since the choice of these variables is arbitrary, such experiments may not provide the required confidence in identified kinetic parameters expressed in terms of standard errors. In this work, an iterative optimization approach is used to determine the above-mentioned variables so as to reduce the experimentation time, while treating the required standard errors as constraints. It is shown using multiple experimental and simulated data that the desired confidence can be reached with much shorter experiments than those generally performed by biosensor users.

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Section: Preliminary Experimental Data and Optimizationsupporting

confidence: 75%

Online optimization of surface plasmon resonance‐based biosensor experiments for improved throughput and confidence

Crescenzo

Woodward

Srinivasan

2008

J of Molecular Recognition

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show abstract

“…Multiplexed platforms have additional problems associated with spot-to-spot variation [32,24] in sensitivity, printed protein concentration and sample depletion where spots upstream in the flow change the concentration of reagents landing on downstream spots. Sample depletion requires mass transport effects to be considered and investigated using non-1:1 model [33] mass transport model. There is no evidence of the breakdown of the 1:1 model given the flow characteristics of our instrument and the number of array spots.…”

Section: Resultsmentioning

confidence: 99%

Label-free Fab and Fc affinity/avidity profiling of the antibody complex half-life for polyclonal and monoclonal efficacy screening

et al. 2015

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A unified approach to affinity screening for Fab and Fc interactions of an antibody for its antigen and FcγR receptor has been developed. An antigen array is used for the Fab affinity and cross-reactivity screening and protein A/G proxy is the FcγR receptor. The affinities are derived using a simple 1:1 binding model with a consistent error analysis. The association and dissociation kinetics are measured over optimised times for accurate determination. The Fab/Fc affinities are derived for ten antibodies: mAb-actin (mouse), pAb-BSA (sheep), pAb-collagen V (rabbit), pAb-CRP (goat), mAb-F1 (mouse), mAbs (mouse) 7.3, 12.3, 29.3, 36.3 and 46.3 raised against LcrV in Yersinia pestis. The rate of the dissociation of antigen-antibody complexes relates directly to their immunological function as does the Fc-FcγR complex and a new half-life plot has been defined with a Fab/Fc half-life range of 17-470 min. The upper half-life value points to surface avidity. Two antibodies that are protective as an immunotherapy define a Fab half-life >250 min and an Fc half-life >50 min as characteristics of ideal interactions which can form the basis of an antibody screen for immunotherapy.

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“…8 With full kinetic/affinity information for biomolecular interactions, we showed that SPR can be used to determine whether the analyte compound binds to one or more sites on the target. High-throughput experiments can be designed by using current SPR array platforms 31,32 for compound/fragment library screening. Compared to common structuralbased techniques (nuclear magnetic resonance, X-ray crystallography), SPR is very cost-effective in terms of protein consumption-only 15 µg of protein is required for achieving suitable immobilization levels and, depending on the protein properties, enabling the screening of hundreds of compounds in a single experiment.…”

Section: Discussionmentioning

confidence: 99%

Biosensor-Based Approach to the Identification of Protein Kinase Ligands with Dual-Site Modes of Action

et al. 2012

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The authors have used a surface plasmon resonance (SPR)-based biosensor approach to identify and characterize compounds with a unique binding mode to protein kinases. Biacore was used to characterize hits from an enzymatic high-throughput screen of the Tec family tyrosine kinase, IL2-inducible T cell kinase (ITK). Complex binding kinetics was observed for some compounds, which led to identification of compounds that bound simultaneously at both the adenosine triphosphate (ATP) binding site and a second, allosteric site on ITK. The presence of the second binding site was confirmed by X-ray crystallography. The second site is located in the N-terminal lobe of the protein kinase catalytic domain, adjacent to but distinct from the ATP site. To enable rapid optimization of binding properties, a competition-based Biacore assay has been developed to successfully identify second site noncompetitive binders that have been confirmed by X-ray crystallographic studies. The authors have found that SPR technology is a key method for rapid identification of compounds with dual-site modes of action.

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Extracting kinetic rate constants from surface plasmon resonance array systems

Cited by 70 publications

References 10 publications

Online optimization of surface plasmon resonance‐based biosensor experiments for improved throughput and confidence

Online optimization of surface plasmon resonance‐based biosensor experiments for improved throughput and confidence

Label-free Fab and Fc affinity/avidity profiling of the antibody complex half-life for polyclonal and monoclonal efficacy screening

Biosensor-Based Approach to the Identification of Protein Kinase Ligands with Dual-Site Modes of Action

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