Direct kinetic assay of interactions between small peptides and immobilized antibodies using a surface plasmon resonance biosensor

Gomes, Paula; Andreu, David

doi:10.1016/s0022-1759(01)00503-8

Cited by 43 publications

(25 citation statements)

References 40 publications

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“…In each sample at 10 kV, there was no observed free αPA FITC peptide, which is typically observed in nonequilibrium capillary electrophoresis of equilibrium mixtures experiments (NECEEM) [28][29][30][31]. The absence of a free peptide peak at nonequilibrium suggests that the peptide is in rapid equilibrium, or rebinding, between the unbound state and the complex with PA, similar to what is often observed in SPR measurements with small molecule analytes or peptides [17].…”

Section: αPa Mobility Shift With Proteinsupporting

confidence: 66%

“…ELISA is used for determining binding dissociation constants (K d ), as well as reagent specificity. SPR can be used for probing association (k on ) and dissociation (k off ) rates along with binding dissociation constants [17][18][19]. Both ELISA and SPR require adsorption or attachment of the peptide or protein target.…”

Section: Introductionmentioning

confidence: 99%

“…The surface attachment or adsorption can impact the molecular recognition by restricting necessary conformational changes or limit binding due to surface steric interactions, as reported with biotin terminated peptides at streptavidin surfaces [20,21]. Alternatively, peptides can be used as the analyte in SPR experiments with immobilized targets, although high concentrations are necessary to achieve the required sensitivity and the risk of binding and rebinding is constantly a problem [17,22].…”

Section: Introductionmentioning

confidence: 99%

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Binding specificity and affinity analysis of an anti-protective antigen peptide reagent using capillary electrophoresis

Kogot¹,

Sarkes²,

Pennington³

et al. 2014

ABB

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Peptide biosensor reagents are emerging as an alternative to typical antibody-based detection methods. Peptides can be rapidly isolated using bacterial display methods for new and emerging biothreats and can be chemically synthesized for rapid, large-scale production. With the emergence of peptide biosensor reagents, there is a growing need to develop methods for characterizing binding interactions. Capillary electrophoresis (CE) is a free-solution separation method that is able to determine target and analyte binding association (K b ) and dissociation constants (K d ). In this study, the K b , K d , and peptide specificity of an isolated peptide binding reagent to protective antigen (PA) of Bacillus anthracis were evaluated using capillary electrophoresis at 10 and 20 kV. The relative binding specificity was rapidly assessed by measuring the peptide relative mobility shift at 20 kV at nonequilibrium using bovine serum albumin (BSA), horseradish peroxidase (HRP), and an anti-PA monoclonal antibody (mAb). The αPA peptide was shown to be highly specific for PA, with a K d = 177 nM measured at 20 kV and K d = 312 nM measured at 10 kV. These results show that peptides from bacterial display libraries can be rapidly tested for specificity and binding affinity, in solution, for use as a potential biosensor reagent against new and emerging biothreats.

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Section: αPa Mobility Shift With Proteinsupporting

confidence: 66%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Binding specificity and affinity analysis of an anti-protective antigen peptide reagent using capillary electrophoresis

Kogot¹,

Sarkes²,

Pennington³

et al. 2014

ABB

View full text Add to dashboard Cite

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“…The great advantages of SPR-based biosensors are that bio-molecular reactions can be monitored in real time, the chip is reusable, and the detection methods enable a simple, rapid, label-free detection [14]. However, its major disadvantage is that it is difficult to determine an analyte at low concentration or with low molecular weight [15]. The advent of dark-field microscope has enabled the study of localized surface plasmon resonance (LSPR) for nanometer-sized metallic structures, which further facilitate its use in label-free detection of various biological analytes in real-time, optical sensing with ultra-high sensitivity [16,17].…”

Section: Introductionmentioning

confidence: 99%

Development of Individual Plasmonic Nanosensors for Clinical Diagnosis

Truong

Sim

2015

IFMBE Proceedings

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“…Mixed monolayers of thiols [an ethanol solution of HS(CH 2 ) 15 COOH (Sigma) and HS(CH 2 ) 11 OH (Sigma) in a ratio of 1 : 300 at an overall concentration of 1 mM] was then deposited over 12 h at 33 ± 1°C. The SMX was immobilized according to the standard procedure [14], i.e., washing with water (5 min), activation of the -COOH groups with a freshly prepared mixture of NHS (0.05 M, Sigma)/EDC (0.2 M, Sigma) (10 min), and exposure in an aqueous solution of SMX with a concentration of 100 mg/mL (10 min). The unreacted active groups were then blocked with ethanolamine (1 M, Sigma) (Fig.…”

mentioning

confidence: 99%

Simple analytical model of biosensor competition analysis for detection of low-molecular-weight analytes

2006

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A steady-state model of competition inhibitor analysis and its application to the detection of sulfamethoxazole in aqueous solutions (0.01-100 mg/mL) using surface plasmon resonance are examined. An analytical expression (logistic curve) relating the equilibrium response of the biosensor to the concentration of the low-molecular-weight analyte, its immobilized analogue, and the biological receptor was obtained.Contemporary advances in the understanding of many biochemical processes and even physiological processes at the molecular level have become possible to a significant degree as a result of successful developments in scientific instrumentation and the creation of direct methods of analysis of intermolecular interactions. Biosensor systems based on optoelectronic and acoustic converters have occupied a special position among such instruments since they make it possible to create devices not requiring additional markers for the investigated molecules, to produce conditions on the surface of the physical converter close to the conditions in biological subjects, to achieve sufficiently high sensitivity for the direct monitoring of the formation of intermolecular complexes, and to secure genuine separation of the intermolecular interaction processes and the production of an informative output signal about the process and its occurrence in real time [1]. Analytical approaches based on surface plasmon resonance converters provide a classical example of biosensor methods for the solution of a wide range of both practical and academic problems. At the same time, in spite of the possibility of detecting surface concentrations of substances right down to picograms per square millimeter, the direct and selective detection of surface concentrations of analytes with molecular masses of less than 5000 Da has come up against serious problems associated with the effect of external factors such as the temperature dependence of the refractive index of a substance (change in the temperature of water by one degree leads to a change in the refractive index by an amount in the order of 10 -4 ) [2]. At the same time analysis of the low-molecular-weight organic xenobiotics such as toxins, antibiotics, pesticides, hormones, etc. is most valuable for practical applications. Competition methods of analysis are in this case a reasonable alternative to direct methods of detection [3][4][5]. It should be noted that competitive methods with immobilized analogues of the analyte and molecular biosystems as selective receptor make it possible to secure an analysis with high operational efficiency as a result of the reproducible formation of a sensitive surface with cross-linked low-molecular-weight organic centers, the long storage period 106 0040-5760/06/4202-0106

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Direct kinetic assay of interactions between small peptides and immobilized antibodies using a surface plasmon resonance biosensor

Cited by 43 publications

References 40 publications

Binding specificity and affinity analysis of an anti-protective antigen peptide reagent using capillary electrophoresis

Binding specificity and affinity analysis of an anti-protective antigen peptide reagent using capillary electrophoresis

Development of Individual Plasmonic Nanosensors for Clinical Diagnosis

Simple analytical model of biosensor competition analysis for detection of low-molecular-weight analytes

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