Infrared up-converting phosphors for bioassays

Corstjens, Paul L. A. M.; Li, S.; Zuiderwijk, Michel; Kardos, Keith; Abrams, William R.; Niedbala, R. Sam; Tanke, Hans J.

doi:10.1049/ip-nbt:20045014

Cited by 152 publications

(183 citation statements)

References 32 publications

Supporting

Mentioning

181

Contrasting

Order By: Relevance

“…1A): 1) 60 min incubation (thermoshaker: 37 °C, 1200 rpm) of 40 µL reporter conjugate with 40 µL of sample; 2) addition of 10 µL (1 ng/µL) of polyclonal biotinylated anti IFN-γ antibody (pB-15-43-07, UCytech, Utrecht, The Netherlands), continued incubation for another 60 min; and, 3) addition of 30 µL LF assay buffer and initiation of LF by addition of the mixture to a microtiterplate well containing a LF strip. The LF strip comprised a sample pad, nitrocellulose membrane with an avidin test line and a rabbit anti-mouse flow control line (12). The assay in summary: a (culture) sample was incubated with an IFN-γ specific reporter to allow binding of IFN-γ to the reporter particle and, upon addition of a biotinylated polyclonal anti IFN-γ antibody an immuno sandwich was formed which could be captured by avidin present on the LF strip.…”

Section: Uligamentioning

confidence: 99%

A user-friendly, highly sensitive assay to detect the IFN-γ secretion by T cells

Corstjens

Zuiderwijk

Tanke

et al. 2008

Clinical Biochemistry

View full text Add to dashboard Cite

Objectives-Development of a user-friendly test alternative to ELISA-based assays to detect IFN-γ by in vitro cultured peripheral blood mononuclear cells (PBMC) stimulated with pathogen-derived antigens.Design and methods-The molecular components of an operational IFN-γ ELISA-based test were applied in a lateral flow (LF) immuno-sandwich assay using up-converting phosphor (UCP) reporter particles. The analytical sensitivity of the UCP-LF IFN-γ assay (ULIGA) was determined and the assay was qualitatively validated with a selection of 60 supernatants derived from PBMC cultures stimulated with M. leprae derived antigens, mitogen or medium alone.Results-ULIGA indicated an analytical sensitivity better than 2 pg/mL, and demonstrated four orders of magnitude dynamic range. The assay correlated well with the IFN-γ ELISA.Conclusions-ULIGA allows detection well below the cutoff value (100 pg/mL) used to define positive responses in the IFN-γ ELISA. The test procedure is less demanding in respect to equipment and labor, and is suited for testing single samples.

show abstract

Section: Uligamentioning

confidence: 99%

A user-friendly, highly sensitive assay to detect the IFN-γ secretion by T cells

Corstjens

Zuiderwijk

Tanke

et al. 2008

Clinical Biochemistry

View full text Add to dashboard Cite

show abstract

“…17 Phosphor emissions can be of different colours, facilitating simultaneous multitarget detection. 32 The UPT particles do not bleach, they facilitate relatively lengthy measurements, and they provide permanent records. 21 UPT particle-based immunochromatographic assays have been reported to have up to 100 times higher sensitivity than conventional fluorescent assays.…”

Section: Introductionmentioning

confidence: 99%

A disposable microfluidic cassette for DNA amplification and detection

et al. 2006

View full text Add to dashboard Cite

; Corstjens, Paul L. A. M.; Mauk, Michael G.; and Bau, Haim H., "A disposable microfluidic cassette for DNA amplification and detection" (2006). Departmental Papers (MEAM). 70. http://repository.upenn.edu/meam_papers/70 A disposable microfluidic cassette for DNA amplification and detection AbstractA pneumatically driven, disposable, microfluidic cassette comprised of a polymerase chain reaction (PCR) thermal cycler, an incubation chamber to label PCR amplicons with upconverting phosphor (UPT) reporter particles, conduits, temperature-activated, normally closed hydrogel valves, and a lateral flow strip, was constructed and tested. The hydrogel valves, which were opened and closed with the aid of electrically controlled thermoelectric units, provided a simple means to seal the PCR reactor and suppress bubble formation. The hydrogel-based flow control was electronically addressable, leakage-free, and biocompatible. To test the device, a solution laden with genomic DNA isolated from B. cereus was introduced into the microfluidic cassette and a specific 305 bp fragment was amplified. The PCR amplicons were labelled with the phosphor (UPT) reporter particles, applied to the lateral flow strip, bound to pre-immobilized ligands, and detected with an IR laser that scanned the lateral flow strip and excited the phosphor (UPT) particles that, in turn, emitted light in the visible spectrum. The UPT particles do not bleach, they provide a permanent record, and they readily facilitate the filtering of background noise. The cassette described herein will be used for rapid testing at the point of care. A pneumatically driven, disposable, microfluidic cassette comprised of a polymerase chain reaction (PCR) thermal cycler, an incubation chamber to label PCR amplicons with upconverting phosphor (UPT) reporter particles, conduits, temperature-activated, normally closed hydrogel valves, and a lateral flow strip, was constructed and tested. The hydrogel valves, which were opened and closed with the aid of electrically controlled thermoelectric units, provided a simple means to seal the PCR reactor and suppress bubble formation. The hydrogel-based flow control was electronically addressable, leakage-free, and biocompatible. To test the device, a solution laden with genomic DNA isolated from B. cereus was introduced into the microfluidic cassette and a specific 305 bp fragment was amplified. The PCR amplicons were labelled with the phosphor (UPT) reporter particles, applied to the lateral flow strip, bound to pre-immobilized ligands, and detected with an IR laser that scanned the lateral flow strip and excited the phosphor (UPT) particles that, in turn, emitted light in the visible spectrum. The UPT particles do not bleach, they provide a permanent record, and they readily facilitate the filtering of background noise. The cassette described herein will be used for rapid testing at the point of care. Comments

show abstract

“…In contrast to conventional fluorescence-based detection, the optical background signal (sometimes referred to as the autofluorescence of the matrix or device material) is virtually zero for UCPs, a distinct advantage when detecting low concentrations of analytes. 12,13,17 Second, UCPs are photochemically stable and do not photobleach or fade. 12,16 Therefore, a phosphor-based assay can be stored indefinitely without a decrease in light-emitting efficiency, can be read by repetitive or time-integrated scans for better counting statistics, and can be archived for subsequent verification.…”

Section: Ucp Technologymentioning

confidence: 99%

“…Corstjens and colleagues, in conjunction with OraSure Technologies, developed a similar portable desktop reader, the Uplink reader, for on-site scanning of captured UCP-labeled biomarkers on lateral-flow test strips. 13 …”

Section: Analytementioning

confidence: 99%