On the mechanism of adsorption of proteins to nitrocellulose in membrane chromatography

Přistoupil, T.I.; Kramlová, M.; Štěrbíková, J.

doi:10.1016/s0021-9673(01)80636-1

Cited by 36 publications

(16 citation statements)

References 13 publications

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“…The capture antibodies in the RZ in the ICR assays for EPX and HNL were more stable in the nitrocellulose membrane if they were first conjugated to PEG, as reported in the literature (14 ). Some of the antibodies had increased activity after the pegylation with a low excess of PEG, possibly from the altered interfacial contacts between the nitrocellulose membrane and the protein.…”

Section: Discussionsupporting

confidence: 55%

Lateral Flow Immunoassay Using Europium (III) Chelate Microparticles and Time-Resolved Fluorescence for Eosinophils and Neutrophils in Whole Blood

Rundström

Jonsson

Mårtensson³

et al. 2007

Clinical Chemistry

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Background:A simple point-of-care method for measuring leukocyte counts in a doctor's office or emergency room could be of great importance. We developed a protocol for measuring cell count by disrupting the cell membrane and analyzing specific proteins within the cells and used it to analyze proteins from eosinophils and neutrophils. Methods: Lateral immunochromatographic (ICR) assays have been developed for eosinophil protein X (EPX) and human neutrophil lipocalin (HNL) as measures of the concentration of eosinophils and neutrophils. The correlation between the lateral ICR assays and cell counting of eosinophils and neutrophils was performed manually and with an automated cell counter. RIA assays measuring the same analytes were also compared with the results from cell counting and lateral ICR assays. Results: The optimized assays showed analytical detection limits below the clinical ranges of 3.36 g/L and 2.05 g/L for EPX and HNL, respectively. The recovery was 114.8%-122.8% for EPX and 94.5%-96.9% for HNL. The imprecision was 3%-17% CV for EPX over the whole range and 5%-16% CV for HNL. The correlation coefficients between manually counted cells and lateral ICR assays were 0.9 and 0.83 for EPX and HNL, respectively.

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Section: Discussionsupporting

confidence: 55%

Lateral Flow Immunoassay Using Europium (III) Chelate Microparticles and Time-Resolved Fluorescence for Eosinophils and Neutrophils in Whole Blood

Rundström

Jonsson

Mårtensson³

et al. 2007

Clinical Chemistry

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“…We hypothesize that this may be due to the negatively charged nature of the nitrocellulose membrane repelling negatively charged GAGs during strip preparation. 87 To check this hypothesis, we have immobilized biotinylated HS15, HEP15, and DEX5 onto the nitrocellulose membrane and flowed streptavidin-coated AuNP. HS15 exhibited a strong band on the nitrocellulose membrane, HEP15 showed a weak band, and for DEX5 no bands were observed ( Figure S19 ).…”

Section: Resultsmentioning

confidence: 99%

GlycoGrip: Cell Surface-Inspired Universal Sensor for Betacoronaviruses

et al. 2021

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Inspired by the role of cell-surface glycoproteins as coreceptors for pathogens, we report the development of GlycoGrip : a glycopolymer-based lateral flow assay for detecting SARS-CoV-2 and its variants. GlycoGrip utilizes glycopolymers for primary capture and antispike antibodies labeled with gold nanoparticles for signal-generating detection. A lock-step integration between experiment and computation has enabled efficient optimization of GlycoGrip test strips which can selectively, sensitively, and rapidly detect SARS-CoV-2 and its variants in biofluids. Employing the power of the glycocalyx in a diagnostic assay has distinct advantages over conventional immunoassays as glycopolymers can bind to antigens in a multivalent capacity and are highly adaptable for mutated strains. As new variants of SARS-CoV-2 are identified, GlycoGrip will serve as a highly reconfigurable biosensor for their detection. Additionally, via extensive ensemble-based docking simulations which incorporate protein and glycan motion, we have elucidated important clues as to how heparan sulfate and other glycocalyx components may bind the spike glycoprotein during SARS-CoV-2 host-cell infection. GlycoGrip is a promising and generalizable alternative to costly, labor-intensive RT-PCR, and we envision it will be broadly useful, including for rural or low-income populations that are historically undertested and under-reported in infection statistics.

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“…In most lateral flow assays, an antibody or antigen specific to the analyte is immobilized onto a nitrocellulose membrane by adsorption forming a test line ( Fig. 2 ) [ 59 , 60 ]. A second zone distal to the point of origin is designated as a control line, where either unbound signaling materials or a second type of particle are captured to indicate that sufficient fluid was applied to indicate successful particle release and capillary flow.…”

Section: Sars-cov-2 Serology Assay Platformsmentioning

confidence: 99%