A Model On-line Flow Injection Fluorescence Immunoassay Using a Protein a Immunoreactor and Lucifer Yellow

Palmer, Diane; Ren, Xuezhen; Fernández-Hernando, P.; Miller, James N.

doi:10.1080/00032719308017972

Cited by 26 publications

(8 citation statements)

References 16 publications

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“…The overall shapes of the curves shown in Figure are similar to those seen in normal competitive binding assays. These curves, and related plots of B / B o versus load A, also show good agreement with those reported in previous studies that have used chromatographic-based simultaneous injection competitive binding immunoassays. ,,,,,,,, The solid lines in Figure show the best-fit response of eq 3 to the experimental data. In obtaining these fits, it was necessary to vary only a single parameter ( S o ) since all other terms in eq 3 (i.e., load A and load A*) had known experimental values.…”

Section: Resultssupporting

confidence: 87%

“…There have been numerous papers in recent years that have used chromatography or flow injection analysis to perform simultaneous injection competitive binding immunoassays. − However, there has been little work concerned with the theoretical basis of this method. In this study, the response and behavior of a simultaneous injection immunoassay will be examined by using chromatographic theory and a model antibody system.…”

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confidence: 99%

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Development of a Theoretical Model for Chromatographic-Based Competitive Binding Immunoassays with Simultaneous Injection of Sample and Label

et al. 1999

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This study examined the theory and behavior of an HPLC-based chromatographic competitive binding immunoassay with the simultaneous injection of sample and a labeled analyte analogue. Equations based on nonlinear chromatographic theory were derived to describe the calibration curve for this assay in a system with adsorption-limited kinetics and homogeneous binding sites. These equations related the assay response (B/Bo) to the column's binding capacity, the moles of injected analyte or labeled analogue, and the flow rate/adsorption kinetics of the system. There was good agreement between the predicted theoretical response and experimental data obtained for the binding of human serum albumin (HSA) to an immobilized anti-HSA antibody column. This theory was also successful in describing the changes that occurred in the calibration curve when the flow rate or amount of labeled analogue applied to the column was varied. A comparison was made between the results of this study and previous theoretical work that examined the behavior of a related, sequential injection competitive binding method. On the basis of the results reported in this work, several general guidelines were developed for the design and optimization of simultaneous injection methods for use in such areas as clinical testing, pharmaceutical analysis, and environmental monitoring.

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Section: Resultssupporting

confidence: 87%

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confidence: 99%

Development of a Theoretical Model for Chromatographic-Based Competitive Binding Immunoassays with Simultaneous Injection of Sample and Label

et al. 1999

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“…The total cycle time between samples was 16 min, and good correlation was noted vs a fluorescence polarization immunoassay (59 ). Other clinical analytes that have been measured by simultaneous injection competitive binding immunoassays include human chorionic gonadotropin (60 ), thyroid-stimulating hormone (60 ), HSA (61 ), IgG (62,63 ), testosterone (64 ), and transferrin (61,65 ); additional studies with theophylline have also been reported (66 -69 ).…”

Section: Competitive Binding Immunoassaysmentioning

confidence: 92%

Affinity Chromatography: A Review of Clinical Applications

Hage

1999

Clinical Chemistry

224

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Affinity chromatography is a type of liquid chromatography that makes use of biological-like interactions for the separation and specific analysis of sample components. This review describes the basic principles of affinity chromatography and examines its use in the testing of clinical samples, with an emphasis on HPLC-based methods. Some traditional applications of this approach include the use of boronate, lectin, protein A or protein G, and immunoaffinity supports for the direct quantification of solutes. Newer techniques that use antibody-based columns for on- or off-line sample extraction are examined in detail, as are methods that use affinity chromatography in combination with other analytical methods, such as reversed-phase liquid chromatography, gas chromatography, and capillary electrophoresis. Indirect analyte detection methods are also described in which immunoaffinity chromatography is used to perform flow-based immunoassays. Other applications that are reviewed include affinity-based chiral separations and the use of affinity chromatography for the study of drug or hormone interactions with binding proteins. Some areas of possible future developments are then considered, such as tandem affinity methods and the use of synthetic dyes, immobilized metal ions, molecular imprints, or aptamers as affinity ligands for clinical analytes.

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“…1), was used as previously described with flow rates of 2-4 ml min 21 and provision for stopped-flow operation. 2 Thiophilic gel reactors of ca. 200 ml volume and an antibody binding capacity (measured using human IgG) of ca.…”

Section: Methodsmentioning

confidence: 99%

Dual analyte flow injection fluorescence immunoassays using thiophilic gel reactors and synchronous scanning detection

Guo

Miller

Evans³

et al. 2000

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Heterogeneous fluorescence immunoassays have been automated using flow injection manifolds incorporating thiophilic gel solid phase reactors to separate antibody-bound and unbound analyte molecules. Antibody elution is achieved by changes in ionic strength, thus allowing the use of pH sensitive fluorescent labels. This facilitates the development of dual analyte systems, in which two competitive immunoassays with separate labels are monitored in parallel. Detection of the fluorophores by high speed synchronous fluorescence scanning while the flow is briefly stopped utilises either one synchronous interval which detects both fluorophores, or two separate scans at different wavelength intervals, one for each fluorophore. Simultaneous analyses of serum albumin and transferrin exemplify these novel approaches. Spectroscopic interferences are very small, analyte recoveries are close to 100%, with a relative standard deviation of 5-6% and a sampling rate of 20 h-1.

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A Model On-line Flow Injection Fluorescence Immunoassay Using a Protein a Immunoreactor and Lucifer Yellow

Cited by 26 publications

References 16 publications

Development of a Theoretical Model for Chromatographic-Based Competitive Binding Immunoassays with Simultaneous Injection of Sample and Label

Development of a Theoretical Model for Chromatographic-Based Competitive Binding Immunoassays with Simultaneous Injection of Sample and Label

Affinity Chromatography: A Review of Clinical Applications

Dual analyte flow injection fluorescence immunoassays using thiophilic gel reactors and synchronous scanning detection

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