Flow-injection enzyme immunoassay of haptens with enhanced chemiluminescence detection

Arefyev, A. A.; Vlasenko, S. B.; Eremin, Sergei A.; Осипов, А. И.; Egorov, A. M.

doi:10.1016/s0003-2670(00)83930-6

Cited by 39 publications

(19 citation statements)

References 5 publications

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“…Even at incubation times up to 5.4 h, a further increase of only 18% was seen versus the results at 2.25 h. Similar results were obtained when utilizing the same amount of antibodies and even larger amounts of HSA (i.e., up to 40 ng/mL). These results agree with previous observations that have been made for one-site immunometric assays involving low-or-intermediate mass targets, for which even shorter incubation times have sometimes been possible [21,27,32]. Based on these results, a minimum incubation time of 30 min and a maximum of 2-2.5 h were used in all later work.…”

Section: Resultssupporting

confidence: 89%

Development of microcolumn-based one-site immunometric assays for protein biomarkers

Pfaunmiller

Anguizola

Milanuk

et al. 2014

Journal of Chromatography A

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One-site immunometric assays that utilize affinity microcolumns were developed and evaluated for the analysis of protein biomarkers. This approach used labeled antibodies that were monitored through on-line fluorescence or near-infrared (NIR) fluorescence detection. Human serum albumin (HSA) was utilized as a model target protein for this approach. In these assays, a fixed amount of labeled anti-HSA antibodies was mixed with samples or standards containing HSA, followed by the injection of this mixture onto an HSA microcolumn to remove excess antibodies and detect the non-retained labeled antibodies that were bound to HSA from the sample. The affinity microcolumns were 2.1 mm i.d. × 5 mm and contained 8-9 nmol of immobilized HSA. These microcolumns were used from 0.10-1.0 mL/min and gave results within 35 s to 2.8 min of sample injection. Limits of detection down to 0.10-0.28 ng/mL (1.5-4.2 pM) or 25-30 pg/mL (0.38-0.45 pM) were achieved when using fluorescein or a NIR fluorescent dye as the label, with an assay precision of ± 0.1-4.2%. Several parameters were examined during the optimization of these assays, and general guidelines and procedures were developed for the extension of this approach for use with other types of affinity microcolumns and protein biomarkers.

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Section: Resultssupporting

confidence: 89%

Development of microcolumn-based one-site immunometric assays for protein biomarkers

Pfaunmiller

Anguizola

Milanuk

et al. 2014

Journal of Chromatography A

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“…There have been numerous papers that have used chromatography and flow-injection analysis to perform simultaneous and sequential injection competitive binding immunoassays (De Alwis and Wilson, 1987;Arefev et al, 1990;Durst et al, 1991;Yap et al, 1991;Pollema et al, 1992a,b;Locascio-Brown et al, 1993;Mecklenburg et al, 1993;Palmer et al, 1993a,b;Pollema and Ruzicka, 1994;Rico et al, 1995;Johns et al, 1996;Martin-Esteban et al, 1996;Valencia-Gonzalez and Diaz-Garcia, 1996;Katmeh et al, 1997;Hage et al, 1993Hage et al, , 1999. However, there have been only a few studies that have examined the theoretical basis of these methods (Hage et al, 1993(Hage et al, , 1999.…”

Section: Introductionmentioning

confidence: 99%

Chromatographic competitive binding immunoassays: a comparison of the sequential and simultaneous injection methods

Nelson

Reiter

Hage

2003

Biomedical Chromatography

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Two approaches for performing competitive binding immunoassays by HPLC and other flow-based systems are the simultaneous and sequential injection methods. Both these techniques make use of a column with a limited amount of antibody, onto which is injected a sample and a fixed amount of a labeled analyte analog. An indirect measure of the unlabeled analyte in the sample is then obtained by looking at the amount of analog in either the nonretained or retained peaks. In the simultaneous injection mode, the sample and labeled analog are applied at the same time to the column, while in the sequential mode the sample is injected first, followed by the analog. This results in a difference in the analytical characteristics of these two approaches. This study used chromatographic theory and previous data obtained for injections of human serum albumin (HSA) onto an anti-HSA antibody column to compare the response, detection limits, range, and sensitivity of these methods. Under equivalent conditions, it was found that the sequential method always provided the best lower limit of detection and sensitivity. However, the simultaneous mode had a broader dynamic range and higher upper limit of detection. From these observations, several guidelines were developed regarding the use and selection of such assays for new applications.

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“…After which, target analyte is passed through the membrane, and the displacement of labeled antigen is monitored downstream. This method has been successfully applied to the detection of numerous hapten molecules and satisfactory sensitivity have been obtained: 10pM for thyroxine (Arefyev et al, 1990); 20 pM for -(difluoromethyl) ornithine (Gunaratna et al, 1993) and 40pM for digoxigenin (Lovgren et al, 1997). The most inviting advantage of this method is the ability to realize automatic detection.…”

Section: Flow Injection Immunoassay (Fiia)mentioning

confidence: 99%

Recent Progress in Noncompetitive Hapten Immunoassays: A Review

Fan¹

2012

Trends in Immunolabelled and Related Techniques

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Flow-injection enzyme immunoassay of haptens with enhanced chemiluminescence detection

Cited by 39 publications

References 5 publications

Development of microcolumn-based one-site immunometric assays for protein biomarkers

Development of microcolumn-based one-site immunometric assays for protein biomarkers

Chromatographic competitive binding immunoassays: a comparison of the sequential and simultaneous injection methods

Recent Progress in Noncompetitive Hapten Immunoassays: A Review

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