Micro-volume wall-less immunoassays using patterned planar plates

Kozak, Katherine R.; Wang, Jianyong; Lye, Melvin; Chuh, Josefa; Takkar, Rashi; Kim, Namyong; Lee, Hyunjae; Jeon, Noo Li; Lin, Kedan; Zhang, Crystal; Wong, Wai Lee; DeForge, Laura

doi:10.1039/c3lc40973b

Cited by 15 publications

(15 citation statements)

References 30 publications

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“…The ATPS-ELISA also provides advantages over miniaturized single ELISA systems, such as Droparray, which use non-aqueous solutions (i.e., oils) to cover the aqueous droplets34. Droparray utilizes droplet sizes (~2 µL) on the same order as the DEX droplets used in our ATPS-ELISA.…”

Section: Discussionmentioning

confidence: 99%

Aqueous two-phase system patterning of detection antibody solutions for cross-reaction-free multiplex ELISA

Frampton

White

Simon

et al. 2014

Sci Rep

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Accurate disease diagnosis, patient stratification and biomarker validation require the analysis of multiple biomarkers. This paper describes cross-reactivity-free multiplexing of enzyme-linked immunosorbent assays (ELISAs) using aqueous two-phase systems (ATPSs) to confine detection antibodies at specific locations in fully aqueous environments. Antibody cross-reactions are eliminated because the detection antibody solutions are co-localized only to corresponding surface-immobilized capture antibody spots. This multiplexing technique is validated using plasma samples from allogeneic bone marrow recipients. Patients with acute graft versus host disease (GVHD), a common and serious condition associated with allogeneic bone marrow transplantation, display higher mean concentrations for four multiplexed biomarkers (HGF, elafin, ST2 and TNFR1) relative to healthy donors and transplant patients without GVHD. The antibody co-localization capability of this technology is particularly useful when using inherently cross-reactive reagents such as polyclonal antibodies, although monoclonal antibody cross-reactivity can also be reduced. Because ATPS-ELISA adapts readily available antibody reagents, plate materials and detection instruments, it should be easily transferable into other research and clinical settings.

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Section: Discussionmentioning

confidence: 99%

Aqueous two-phase system patterning of detection antibody solutions for cross-reaction-free multiplex ELISA

Frampton

White

Simon

et al. 2014

Sci Rep

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“…A more detailed description about fabrication of DropArray can be found elsewere. 12,13 Each circular plastic area is rendered hydrophilic with a plasma treatment. Each circular well can then accommodate a drop of 2-4 mL in volume (Fig.…”

Section: Droparray Plate and Droparray Washermentioning

confidence: 99%

“…A more detailed description about DropArray washing station properties and theory are available elsewhere. [12][13][14] Cell Seeding and Compound Treatment on the DropArray Plate HeLa (ATCC) were cultured in Nunc tissue culture flask in D10 growth media (Dulbecco's modified Eagle's medium [DMEM] high glucose [4.5 g/L] supplemented with 10% fetal calf serum, 5% penicillin-streptomycin). The cells were maintained in a humidified 37°C incubator with an air to 5% CO 2 atmosphere.…”

Section: Droparray Plate and Droparray Washermentioning

confidence: 99%

“…DropArray also offers a different washing method using laminar flow, which circumvents the classic microwell-based washing limitations by allowing gentle but efficient washing with high cell retention. 12,13 In this study, the use of the DropArray technology enabled the development of a mitotic index assay, which was validated with the antimitotic drug docetaxel.…”

Section: Introductionmentioning

confidence: 99%

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Miniaturization of Mitotic Index Cell-Based Assay Using “Wall-Less” Plate Technology

Guezennec¹,

Phong²,

Nor³

et al. 2014

ASSAY and Drug Development Technologies

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The use of microscopic imaging for the accurate assessment of cells in mitosis is hampered by the round morphology of mitotic cells, which renders them poorly adherent and highly susceptible to loss during the washing stage of cell-based assays. Here, to circumvent these limitations, we make use of DropArray, a recent technology that allows high retention of weakly adherent cells and suspension cells. DropArray offers the competitive advantage of maintaining the classic high throughput format of microtiter plates while reducing classic microwell volume by up to 90% by using a drop format. Here, we present a mitotic index cell-based assay using the mitosis marker phospho histone H3 at serine 10 on a DropArray 384-well plate format. Dose-response curve analysis of the mitotic index assay with an antimitotic drug (docetaxel) on DropArray is presented that shows an effective dosage compared to previous established results similar to those obtained with conventional microtiter plates. The mitotic index assay with DropArray showed a Z-factor >0.6. Our results validate DropArray as a suitable platform for high throughput screening for compounds affecting mitosis or the cell cycle.

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“…Moreover, it enables many-fold reduction in reagent costs by using minimal amount of reagents, and has significantly reduced the number of steps in comparison to conventional ELISA. Similarly, various ultrasensitive microfluidic ELISA formats have also been demonstrated recently by other researchers [10,[16][17][18][19]. These developments have enabled the researchers to proceed towards lab-on-a-chip technologies, which will be the main area of immunodiagnostics research in the near future.…”

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confidence: 99%

Immunodiagnostics: Major Advances and Future Insights

Vashist¹

2013

J Biochip Tissue Chip

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There have been tremendous advances in immunodiagnostics during the last five decades, which have led to the emergence of potential diagnostic technologies, new immunoassay formats and diagnostic platforms. The radioimmunoassay and Enzyme Linked Immunosorbent Assay (ELISA) were the predominant techniques during the initial era of immunodiagnostics. The strenuous research efforts during the last three decades have enabled the development of potential antibodies for a wide range of analytes. Till date,t ELISA has been the gold standard of clinical immunodiagnostics, which has also been used very frequently for industrial analysis, environmental monitoring, food analysis and bioanalytical sciences. Despite the numerous advances in the field, it is difficult for any other technique to achieve the same high sensitivity, specificity, precision and throughput as ELISA. Various robotic work stations have been devised to perform automated ELISAs in high volume settings, such as in hospitals and industry. Similarly, the Microtiter Plate (MTP) readers have been improved significantly in terms of optics, technology and data analysis. In addition to conventional 96-well MTP format, various high throughput formats, such as 384 and 1536 wells, have also been devised for specific needs. The last two decades have seen considerable improvements in antibody immobilization strategies [1][2][3], which have significantly improved the analytical performance of ELISA in terms of higher sensitivity, greater detection range, lower limit of detection and reduced assay duration [4][5][6]. Similarly, the sample preparation strategies and screening of immunological components have also been critically improved, which enables highly reproducible immunoassays [7,8]. The development of highly sensitive enzyme substrates have led to ultrasensitive ELISAs and chemiluminescent immunoassays [9], while the use of superior fluorescent dyes improved the fluorescent immunoassays. Additionally, the use of infra-red dyes, quantum dots, gold nanoparticles, beads, and other nanomaterial/nanocomposites enabled the development of new ELISA formats, such as the naked-eye ELISA that was demonstrated using gold nanoparticles [10].The development of bead-based AlphaLISA by Perkin Elmer [11] is another major development in immunodiagnostics as it has critically reduced the immunoassay duration and complexity. It employs a streptavidin-coated Alpha donor bead and anti-analyte-conjugated AlphaLISA acceptor bead, which come into close proximity to generate the signal in the presence of analyte. These chemiluminescent immunoassays do not employ any washing step and have high sensitivity, greater dynamic range and superior analytical perfromance than the conventional ELISA.The continuous developments in microfluidic technologies have critically reduced the assay duration and number of process steps apart from enabling the automated handling of process steps. The emergence of surface plasmon resonance (SPR)-based real-time and label-free immunoassays using the Biacore ...

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Micro-volume wall-less immunoassays using patterned planar plates

Cited by 15 publications

References 30 publications

Aqueous two-phase system patterning of detection antibody solutions for cross-reaction-free multiplex ELISA

Aqueous two-phase system patterning of detection antibody solutions for cross-reaction-free multiplex ELISA

Miniaturization of Mitotic Index Cell-Based Assay Using “Wall-Less” Plate Technology

Immunodiagnostics: Major Advances and Future Insights

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