Convection during Incubation of Microplate Solid Phase Immunoassay:  Effects on Assay Response and Variation

Beumer, Tom; Haarbosch, Paul; Carpay, Wim

doi:10.1021/ac950696s

Cited by 17 publications

(7 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This by far exceeds the maximum heat flux that can be provided by the heating element, which makes heat transport by thermal convection even more likely. The currently found thermal gradients are a factor 3−5 smaller than those predicted from the scaled-up experiments . Even though mixing boundary layers must therefore be slightly thicker than predicted in, calculations on immunoassay incubation predict only marginal effects to reaction kinetics.…”

Section: Resultsmentioning

confidence: 63%

“…The currently found thermal gradients are a factor 3−5 smaller than those predicted from the scaled-up experiments . Even though mixing boundary layers must therefore be slightly thicker than predicted in, calculations on immunoassay incubation predict only marginal effects to reaction kinetics. A more considerable deviation from our earlier data is the finding of a second high-temperature region close to the liquid surface.…”

Section: Resultsmentioning

confidence: 63%

“…A more considerable deviation from our earlier data is the finding of a second high-temperature region close to the liquid surface. One probable cause for these deviations is the erroneous use of scaling parameters in ref .

2 Reconstruction of the temperature distribution T ( x,y,z ) in the central plane of symmetry of the cuvet according to Abel's method in equilibrated temperature situation.…”

Section: Resultsmentioning

confidence: 99%

“…This heating does provide significantly higher signal levels. Recently we demonstrated that the increase is only to a limited extent due to the assumed improved association constant between reactants. Far more important is the thermal mixing that enhances the diffusion-limited reaction at low concentration levels.…”

mentioning

confidence: 97%

See 3 more Smart Citations

Nonintrusively Measured Temperature Distributions as Evidence for Free Convection in Immunoassay Incubation

Beumer¹,

Timmerman²

1997

Anal. Chem.

Self Cite

View full text Add to dashboard Cite

Immunoassay incubation is classically performed at elevated temperatures to speed up reaction processes. This acceleration has long been assumed to be due to an increased association constant between reactants. Using an interferometric method to visualize temperature differences inside a small reaction cuvet, we demonstrate that the temperature profiles inside the liquid cannot be caused by conduction but only by thermal convection. Numerical simulations further support this experimental evidence. This paper conclusively demonstrates that thermally induced mass transport occurs in immunoassay incubation. Current data show that earlier estimates of heat transport coefficients in such incubations have been probably overestimated by a factor 3.

show abstract

Section: Resultsmentioning

confidence: 63%

Section: Resultsmentioning

confidence: 63%

2 Reconstruction of the temperature distribution T ( x,y,z ) in the central plane of symmetry of the cuvet according to Abel's method in equilibrated temperature situation.…”

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 97%

See 2 more Smart Citations

Nonintrusively Measured Temperature Distributions as Evidence for Free Convection in Immunoassay Incubation

Beumer¹,

Timmerman²

1997

Anal. Chem.

Self Cite

View full text Add to dashboard Cite

show abstract

“…A major difficulty with immunoassays is assay variation attributed to both microplate-and nonmicroplate-associated factors, and compounded by a complicity of these factors leading to exacerbation of assay variability. Operator competency; microplate, calibrator, and reagent lot differences; microplate edge effects arising from temperature and evaporation rate differentials; and convective forces during assay incubation and washing steps have been reported to contribute to variations in well-to-well and plateto-plate signal read out (1)(2)(3)(4)(5)(6)(7)(8). Microplate variability with row and/or column effects within the microplate as well as systematic differences between microplates has been observed (7,8).…”

mentioning

confidence: 99%

Variability in Microplate Surface Properties and Its Impact on ELISA

Lilyanna

Moriguchi

et al. 2018

The Journal of Applied Laboratory Medicine

View full text Add to dashboard Cite

Background: Microplate-based immunoassays are widely used in clinical and research settings to measure a broad range of biomarkers present in complex matrices. Assay variability within and between microplates can give rise to false-negative and false-positive results leading to incorrect conclusions. To date, the contribution of microplates to this variability remains poorly characterized and described. This study provides new insights into variability in immunoassays attributable to surface characteristics of commercial microplates. Methods: Well-to-well assay variation in γ-treated and nontreated 96-well opaque microplates suitable for chemiluminescence assays was determined by use of a validated sandwich ELISA. Microplate surface characteristics were assessed by sessile drop contact angle measurements, scanning electron microscopy, energy dispersive x-ray spectroscopy, and atomic force microscopy. Results: All microplate types tested exhibited vendor-specific assay response profiles; and "rogue" plates with very high intraassay variation and deviant mean assay responses were found. Within-plate, location-dependent bias in assay responses and variability in well contact angle were also observed. We demonstrate substantial differences in well-surface properties with putative effects on protein-coating reproducibility and hence consistency in immunoassay responses. A surface "cleaning" effect on manufacturing residues was attributed to γirradiation, and treated microplates manifest increased polar functionalities, surface roughness, and assay responses. Conclusions: Our data suggest that tighter control of variability in surface roughness, wettability, chemistry, and level of residual contaminants during microplate preparation is warranted to improve consistency of ELISA assay read out. IMPACT STATEMENT In clinical and research settings, microplate-based immunoassays are subject to assay variability within and between microplates that can give rise to false-negative and false-positive results leading to incorrect conclusions. This article clarifies the contribution of microplates to assay variability, which is poorly understood to date. Detailed physicochemical analyses of microplate well surfaces provided new insights into immunoassay variability attributable to microplate surface properties and preparation processes. Our work prompts further investigations to elucidate surface parameters that influence antibody adsorption and biofunctionality, highlighting manufacturing issues that require further review to overcome problems with microplate variability and assay performance.

show abstract

Homogeneous Two-Site Immunometric Assay Kinetics as a Theoretical Tool for Data Analysis

Zuber

Mathis²,

Flandrois

1997

Analytical Biochemistry

View full text Add to dashboard Cite

Convection during Incubation of Microplate Solid Phase Immunoassay: Effects on Assay Response and Variation

Cited by 17 publications

References 19 publications

Nonintrusively Measured Temperature Distributions as Evidence for Free Convection in Immunoassay Incubation

Nonintrusively Measured Temperature Distributions as Evidence for Free Convection in Immunoassay Incubation

Variability in Microplate Surface Properties and Its Impact on ELISA

Homogeneous Two-Site Immunometric Assay Kinetics as a Theoretical Tool for Data Analysis

Contact Info

Product

Resources

About