Critical Comparison between Large and Mini Vertical Flow Immunoassay Platforms for <i>Yersinia Pestis</i> Detection

Devadhasan, Jasmine Pramila; Gu, Jian; Chen, Peng; Smith, S. D.; Thomas, Baiju; Gates-Hollingsworth, Marcellene A.; Hau, Derrick; Pandit, Sujata G.; AuCoin, David P.; Zenhausern, Frédéric

doi:10.1021/acs.analchem.0c05278

Cited by 19 publications

(14 citation statements)

References 39 publications

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“…Furthermore, the top mAb pair for detecting LcrV by LFI was evaluated in a vertical flow immunoassay (VFI) format. The mini VFI prototype can detect as low as 25 pg/mL LcrV [ 67 ].…”

Section: Discussionmentioning

confidence: 99%

Development of a dual antigen lateral flow immunoassay for detecting Yersinia pestis

et al. 2022

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Background Yersinia pestis is the causative agent of plague, a zoonosis associated with small mammals. Plague is a severe disease, especially in the pneumonic and septicemic forms, where fatality rates approach 100% if left untreated. The bacterium is primarily transmitted via flea bite or through direct contact with an infected host. The 2017 plague outbreak in Madagascar resulted in more than 2,400 cases and was highlighted by an increased number of pneumonic infections. Standard diagnostics for plague include laboratory-based assays such as bacterial culture and serology, which are inadequate for administering immediate patient care for pneumonic and septicemic plague. Principal findings The goal of this study was to develop a sensitive rapid plague prototype that can detect all virulent strains of Y. pestis. Monoclonal antibodies (mAbs) were produced against two Y. pestis antigens, low-calcium response V (LcrV) and capsular fraction-1 (F1), and prototype lateral flow immunoassays (LFI) and enzyme-linked immunosorbent assays (ELISA) were constructed. The LFIs developed for the detection of LcrV and F1 had limits of detection (LOD) of roughly 1–2 ng/mL in surrogate clinical samples (antigens spiked into normal human sera). The optimized antigen-capture ELISAs produced LODs of 74 pg/mL for LcrV and 61 pg/mL for F1 when these antigens were spiked into buffer. A dual antigen LFI prototype comprised of two test lines was evaluated for the detection of both antigens in Y. pestis lysates. The dual format was also evaluated for specificity using a small panel of clinical near-neighbors and other Tier 1 bacterial Select Agents. Conclusions LcrV is expressed by all virulent Y. pestis strains, but homologs produced by other Yersinia species can confound assay specificity. F1 is specific to Y. pestis but is not expressed by all virulent strains. Utilizing highly reactive mAbs, a dual-antigen detection (multiplexed) LFI was developed to capitalize on the diagnostic strengths of each target.

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“…Furthermore, the top mAb pair for detecting LcrV by LFI was evaluated in a vertical flow immunoassay (VFI) format. The mini VFI prototype can detect as low as 25 pg/mL LcrV [ 67 ].…”

Section: Discussionmentioning

confidence: 99%

Development of a dual antigen lateral flow immunoassay for detecting Yersinia pestis

et al. 2022

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“…After drying, the VFI membrane disks were scanned with a flatbed optical scanner (CanoScan 9000 FII) to measure the signal intensity of the test spots as described in previous literature. 1 …”

Section: Materials and Methodsmentioning

confidence: 99%

“…Samples were flown through a custom-built VFI device at 0.2 mL/min and dried for 10 min before scanning. After drying, the VFI membrane disks were scanned with a flatbed optical scanner (CanoScan 9000 FII) to measure the signal intensity of the test spots as described in previous literature …”

Section: Materials and Methodsmentioning

confidence: 99%

“…Antibody microarrays are currently being used for numerous applications including analysis of proteins, nucleic acids, cell surface proteins, and simultaneous detection of these analytes. − They have contributed significantly to “life-omics”, as well as disease diagnostics and management . In general, antibody arrays are fabricated by immobilizing multiple antibodies on a solid substrate, such as a chemically treated glass slide or well plate, or a nitrocellulose (NC) paper membrane, to analyze antibody–antigen interactions .…”

Section: Introductionmentioning

confidence: 99%

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Optimization of an Antibody Microarray Printing Process Using a Designed Experiment

Devadhasan

et al. 2022

ACS Omega

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Antibody microarrays have proven useful in immunoassay-based point-of-care diagnostics for infectious diseases. Noncontact piezoelectric inkjet printing has advantages to print antibody microarrays on nitrocellulose substrates for this application due to its compatibility with sensitive solutions and substrates, simple droplet control, and potential for high-capacity printing. However, there remain real-world challenges in printing such microarrays, which motivated this study. The effects of three concentrations of capture antibody (cAb) reagents and nozzle hydrostatic pressures were chosen to investigate three responses: the number of printed membrane disks, dispensing performance, and microarray quality. Printing conditions were found to be most ideal with 5 mg/mL cAb and a nozzle hydrostatic pressure near zero, which produced 130 membrane disks in a single print versus the 10 membrane disks per print before optimization. These results serve to inform efficient printing of antibody microarrays on nitrocellulose membranes for rapid immunoassay-based detection of infectious diseases and beyond.

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“…Nevertheless, although the cross-reactivity of the analyte may lead to negligible influence on two target detections, this might be a problem for further consideration in the tests of more kinds of targets. Fortunately, for a model like multiplex vertical flow immunoassay, 45 which avoids the spatial location differences between different captured antigens, analyte cross-reactivity would not be exerted in the CNAN system.…”

Section: ■ Conclusionmentioning

confidence: 99%

Self-Assembling Antibody Network Simplified Competitive Multiplex Lateral Flow Immunoassay for Point-of-Care Tests

Zhou

et al. 2022

Anal. Chem.

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Multiplex lateral flow immunoassay (mLFIA) has attracted great attention due to the increasing need for rapid detection of multiple analytes. However, it has a number of disadvantages with regard to accuracy and interference because of difficulties in simplifying the process of preparing nanomaterial-based probes. In this work, inspired by protein self-assembly, for the first time, a facile natural antibody network (NAN)-based mLFIA for multiple chloramphenicol (CAP) and streptomycin (STR) determination was designed. The NAN structure was constructed by introducing a second antibody (Ab2) as a scaffold to noncovalently combine with various monoclonal antibodies (mAbs), thus permitting each mAb to act as an independent functional unit to maintain bioactivity. Furthermore, the NAN was colored by simple one-step staining using coomassie brilliant blue R-250 (CBBR) to form a chromogenic probe, eliminating the need for complex nanomaterials to improve reproducibility and precision. Under optimal conditions, a satisfactory detection performance (the visual limit of detection (v-LOD) of 3 ng mL–1 for CAP and 20 ng mL–1 for STR) was obtained for whole milk analysis, which met the basic requirement of detection and had good specificity, reproducibility (relative standard deviation (RSD) < 15%), and robustness. In addition, the precision of the detection results was improved usefully since the test procedure was simplified. Overall, the developed system enables fast, simple, and reliable point-of-care assays of multiple analytes.

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Critical Comparison between Large and Mini Vertical Flow Immunoassay Platforms for Yersinia Pestis Detection

Cited by 19 publications

References 39 publications

Development of a dual antigen lateral flow immunoassay for detecting Yersinia pestis

Development of a dual antigen lateral flow immunoassay for detecting Yersinia pestis

Optimization of an Antibody Microarray Printing Process Using a Designed Experiment

Self-Assembling Antibody Network Simplified Competitive Multiplex Lateral Flow Immunoassay for Point-of-Care Tests

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