The Immunoprobe Aggregation State is Central to Dipstick Immunoassay Performance

Hristov, Delyan R.; Pimentel, Alyssa Jean; Ujialele, Godwin; Hamad‐Schifferli, Kimberly

doi:10.1021/acsami.0c08628

Cited by 19 publications

(29 citation statements)

References 47 publications

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“…Immunochromatographic assays (ICAs) are the workhorses of point-of-care testing (POCT) for monitoring of analytes, such as viruses, cancer biomarkers, bacteria, , and mytoxins, at low concentrations, particularly playing a significant role in the COVID-19 pandemic . The analytical power of such techniques benefits from the strong bioactivity of monoclonal antibodies (mAbs) that can recognize targets and the aggregation-induced optical signals of nanomaterials (NMs) . Recently, myriad signal-marking materials, comprising polymer dots, Ru(bpy) 3 2+ -loaded mesoporous silica nanoparticles, magnetic fluorescent beads, Pt–Ni(OH) 2 nanosheets, perovskite nanocrystals, and microorganisms (MOs)@Au, , have been employed to lower the detection limit in ICAs.…”

mentioning

confidence: 99%

Diverse Dyes-Embedded Staphylococcus aureus as Potential Biocarriers for Enhancing Sensitivity in Biosensing

Zhao

Bai

et al. 2021

Anal. Chem.

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Nanomaterials-based immunochromatographic assays (ICAs) have gained great commercial success in real-life point-of-care testing (POCT). Exploring novel carriers of ICAs with improved signaling and sustained activity favors the development of sensitive POCT. Herein a potent signal biotag, colored Staphylococcus aureus (SA), was created for ICA carriers through a mild self-assembly strategy, providing high luminance and abundant specific binding sites for immobilization of monoclonal antibodies (mAbs). The biocompatible SA-dyes (SADs) retained both an intact surface structure for mAbs labeling (Fc portion) and the superior bioactivity of immobilized mAbs (affinity constant was about 109 M–1), thus waiving the intrinsic limitations of traditional nanomaterials and endowing high sensitivity. Proof-of-concept was demonstrated by employing Congo red- or/and fluorescein isothiocyanate-embedded SA (SACR, SAFITC, and SACR–SAFITC) as ICA carriers to detect zearalenone (ZEN) through colorimetric or/and fluorimetric signals. Furthermore, the ICAs satisfied the clinical requirement perfectly, including limit of detection (0.013 ng/mL, which was at least an 85-fold improvement over that of traditional gold nanoparticles-based ICA), linearity (R 2 > 0.98), reproducibility (RSD < 8%), selectivity, and stability. Importantly, the proposed biosensors could be well-applied in four real samples for ZEN monitoring with satisfactory recoveries, correlating well with the results from liquid chromatography–tandem mass spectrometry (LC-MS). This work also proved a universal design for tailoring coloration bands for SAD–ICA detection of multiple analytes.

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mentioning

confidence: 99%

Diverse Dyes-Embedded Staphylococcus aureus as Potential Biocarriers for Enhancing Sensitivity in Biosensing

Zhao

Bai

et al. 2021

Anal. Chem.

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“…Prior studies have shown that the protein corona formed around the NP-Ab conjugate from the proteins in the patient sample can impact sandwich immunoassay formation with the target 27 and that the immunoassay performance is sensitive to NP aggregation. 16 Therefore, the sample matrix can impact the performance. BSA was used here as a test case in which the performance could be quantified and compared to a behavior in the running media of human saliva and serum, which were investigated as real biological media.…”

Section: Discussionmentioning

confidence: 99%

“…Immunoprobe synthesis followed our previous work. 16 An appropriate amount of as-synthesized NPs was pipetted into a LoBind Eppendorf (Sigma). A 1 mg/mL solution (6.5 μM) of the antibody (10 μL) ( Table S2 ) was added per mL of AuNPs for a final antibody concentration of 0.065 μM.…”

Section: Methodsmentioning

confidence: 99%

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Developing a Paper-Based Antigen Assay to Differentiate between Coronaviruses and SARS-CoV-2 Spike Variants

et al. 2021

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COVID-19 first appeared in December of 2019 in Wuhan, China. Since then, it has become a global pandemic. A robust and scalable diagnostics strategy is crucial for containing and monitoring the pandemic. RT-PCR is a known, reliable method for COVID-19 diagnostics, which can differentiate between SARS-CoV-2 and other viruses. However, PCR is location-dependent, time-consuming, and relatively expensive. Thus, there is a need for a more flexible method, which may be produced in an off-the-shelf format and distributed more widely. Paper-based immunoassays can fulfill this function. Here, we present the first steps toward a paper-based test, which can differentiate between different spike proteins of various coronaviruses, SARS-CoV-1, SARS-CoV-2, and CoV-HKU1, with negligible cross-reactivity for HCoV-OC43 and HCoV-229E in a single assay, which takes less than 30 min. Furthermore, our test can distinguish between fractions of the same spike protein. This is done by an altered assay design with four test line locations where each antigen builds a unique, identifiable binding pattern. The effect of several factors, such as running media, immunoprobe concentration, and antigen interference, is considered. We find that running media has a significant effect on the final binding pattern where human saliva provides results while human serum leads to the lowest signal quality.

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“…Synthesis was done in accordance with the principles laid out in our previous work. 3 The appropriate amount of as synthesised AuNPs were pipetted into a LoBind Eppendorf (Sigma, product code: Z66613). The choice of vessel was deliberate as other plastics led to NP sticking to the sides.…”

Section: Immunoprobe Synthesismentioning

confidence: 99%

Developing a Paper-Based Antigen Assay to Differentiate Between Coronaviruses and SARS-CoV-2 Spike Variants

Hristov¹,

Rijal²,

Gómez-Márquez³

et al. 2020

Preprint

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COVID-19 first appeared in December of 2019 in Wuhan, China. Since then it has become a global pandemic. A robust and scalable diagnostics strategy is crucial for containing and monitoring the pandemic. RT-PCR is a known, reliable method for COVID-19 diagnostics which can differentiate between SARS-CoV-2 and other viruses. However, PCR is location dependent, time consuming and relatively expensive. Thus, there is a need for a more flexible method which may be produced in an off-the-shelf format and distributed more widely. Paper-based immunoassays can fulfill this function. Here we present the first steps towards a paper-based test which can differentiate between different between the Spike protein of various coronaviruses, SARS-CoV-1, SARS-CoV-2 and CoV-HKU1 with negligible cross reactivity for HCoV-OC43 and HCoV-229E in a single assay which takes less than 30 minutes. Furthermore, our test can distinguish between fractions of the same Spike protein. This is done by an altered assay design with four test line locations where each antigen builds a unique, identifiable binding pattern. The effect of several factors, such as running media, immunoprobe concentration and antigen interference is considered. We find that running media has a significant effect on the final binding pattern where human saliva provides results while human serum leads to the lowest signal quality. <br>

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The Immunoprobe Aggregation State is Central to Dipstick Immunoassay Performance

Cited by 19 publications

References 47 publications

Diverse Dyes-Embedded Staphylococcus aureus as Potential Biocarriers for Enhancing Sensitivity in Biosensing

Diverse Dyes-Embedded Staphylococcus aureus as Potential Biocarriers for Enhancing Sensitivity in Biosensing

Developing a Paper-Based Antigen Assay to Differentiate between Coronaviruses and SARS-CoV-2 Spike Variants

Developing a Paper-Based Antigen Assay to Differentiate Between Coronaviruses and SARS-CoV-2 Spike Variants

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