Surface Plasmon Resonance Detection and Multispot Sensing for Direct Monitoring of Interactions Involving Low-Molecular-Weight Analytes and for Determination of Low Affinities

Karlsson, Robert; Stahlberg, R.

doi:10.1006/abio.1995.1350

Cited by 268 publications

(139 citation statements)

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“…We analyzed the affinities of our ligands using referencesubtracted response levels at equilibrium to determine the binding isotherm (Figure 3) (34). In the case of low-affinity ligands, we were able to obtain approximate fits using a single-site model (eq 1) by assuming the curves would reach a similar plateau.…”

Section: Resultsmentioning

confidence: 99%

Affinity-Based Inhibition of β-Amyloid Toxicity

et al. 2002

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Strategies for interfering with protein aggregation are important for elucidating and controlling the pathologies of amyloid diseases. We have previously identified compounds that block the cellular toxicity of the -amyloid peptide, but the relationship between their ability to inhibit toxicity and their affinity for A is unknown. To elucidate this relationship, we have developed an assay capable of measuring the affinities of small molecules for -amyloid peptide. Our approach employs immobilized -amyloid peptide at low density to minimize the problems that arise from variability in the -amyloid aggregation state. We found that low-molecular weight (MW of 700-1700) ligands for -amyloid can be identified readily by using surface plasmon resonance. The best of these bound effectively (K d ∼ 40 µM) to -amyloid. The affinities measured for peptides in the SPR assay correspond to results from A cell toxicity assays. The most potent ligands for immobilized -amyloid are the most potent inhibitors of the neuronal cell toxicity of -amyloid. Compounds with dissocation constants above ∼100 µΜ did not show significant activity in the cell toxicity assays. Our data support the hypothesis that ligands exhibiting greater affinity for the -amyloid peptide are effective at altering its aggregation and inhibiting cell toxicity.

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Section: Resultsmentioning

confidence: 99%

Affinity-Based Inhibition of β-Amyloid Toxicity

et al. 2002

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“…After each run, regeneration of the sensor chip surface was accomplished by two successive injections of 15 l of 5 mM HCl. The values for rate constants were determined by nonlinear regression analyses using BIAevaluation 2.1 software provided by the manufacturer and as described in detail by Karlsson et al (32,33). Association rate constants (k a ) were calculated from the linear portion of sensorgrams during the early association phase.…”

Section: Methodsmentioning

confidence: 99%

Identification of Hyaluronan-binding Domains of Aggrecan

Watanabe

Cheung

Itano

et al. 1997

Journal of Biological Chemistry

126

102

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“…However, given the large number of variant peptides, the only practical way to perform the screening consists of monoclonal antibody (MAb) immobilization and injection of peptide analytes. As direct detection of analytes smaller than 5000 Da is usually unfeasible with standard BIAcore protocols [20,21], we have carried out an extensive optimization of this approach using 44 of the 240 A15 single mutants as analytes and anti-site A MAb SD6 as ligand. The generally good agreement found between biosensor data and the anities known from ELISA [18] validates our analytical method.…”

Section: Introductionmentioning

confidence: 99%

Surface plasmon resonance screening of synthetic peptides mimicking the immunodominant region of C-S8c1 foot-and-mouth disease virus

Gomes

Giralt

Andreu

1999

Vaccine

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The main antigenic site (site A) of foot-and-mouth disease virus (FMDV, strain C-S8c1) may be adequately reproduced by a 15-peptide with the amino acid sequence H-YTASARGDLAHLTTT-NH 2 (A15), corresponding to the residues 136±150 of the viral protein VP1. The eect of amino acid substitutions within A15 on its antigenicity towards monoclonal antibodies (MAb) raised against antigenic site A, has been studied by means of BIAcore technology, based on surface plasmon resonance (SPR). Although these antigenicities have previously been determined from enzyme-linked immunosorbent assays (ELISA), the SPRbased technique is superior in that it allows a fast and straightforward screening of antigens while simultaneously providing kinetic data of the antigen±antibody interaction. With a view to screening fairly large libraries of individual peptides, we have inverted the typical SPR experiment by immobilizing the MAb on the sensor surface and using peptides as soluble analytes. We report the validation of this approach through the screening of 44 site A peptides, with results generally in good agreement with the relative antigenicities previously determined by competition ELISA. #

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Surface Plasmon Resonance Detection and Multispot Sensing for Direct Monitoring of Interactions Involving Low-Molecular-Weight Analytes and for Determination of Low Affinities

Cited by 268 publications

References 0 publications

Affinity-Based Inhibition of β-Amyloid Toxicity

Affinity-Based Inhibition of β-Amyloid Toxicity

Identification of Hyaluronan-binding Domains of Aggrecan

Surface plasmon resonance screening of synthetic peptides mimicking the immunodominant region of C-S8c1 foot-and-mouth disease virus

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