Prototyping of a lateral flow assay based on monoclonal antibodies for detection of Bothrops venoms

Knudsen, Carsten; Jürgensen, Jonas A.; Knudsen, Pelle D.; Oganesyan, Irina; Harrison, Julian A.; Dam, Søren H.; Haack, Aleksander M.; Friis, Rasmus U.W.; Vitved, Lars; Belfakir, Selma B.; Ross, G.M.S.; Zenobi, Renato; Laustsen, Andreas Hougaard

doi:10.1016/j.aca.2023.341306

Cited by 4 publications

(8 citation statements)

References 31 publications

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“…The LOD value was found to be 375 ng mL −1 using the equation LOD = 3.3( σ / S ), where σ is the standard deviation of the test line signal, S is the slope of the curve, and 3.3 is a constant. 26,27 For every concentration, three trials were conducted, followed by the calculation of their average. The LOD was calculated as the mean plus three times the standard deviation of the no-analyte control LFAs.…”

Section: Resultsmentioning

confidence: 99%

A novel lateral flow immunochromatographic assay using a recombinant VP2 antigen for total antibody detection of canine parvovirus-2

Salmanli,

Tezcan,

Karaoglu

2024

Anal. Methods

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Section: Resultsmentioning

confidence: 99%

A novel lateral flow immunochromatographic assay using a recombinant VP2 antigen for total antibody detection of canine parvovirus-2

Salmanli,

Tezcan,

Karaoglu

2024

Anal. Methods

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“…The Lachesis -mAb sandwich pair were discovered using the method previously reported for the Bothrops- mAb pair 38 . All protocols in this study were approved by the Research Ethical Committee of University of Southern Denmark, following the guidelines of the Danish National Animal Ethics Committee (J-Number 2015-15-0201-00680), and were performed according to the ARRIVE guidelines.…”

Section: Methodsmentioning

confidence: 99%

“…5 ng/mL (with enrichment) in plasma for N. atra Spiked human plasma 31 Singleplex B. multicinctus Asia Rabbit pAbs AuNPs 10–15 min 1 ng/mL in buffer for B. multicinctus Tissue homogenates, blood, and urine from envenomed rats 37 Singleplex Bothrops spp. Latin America Mouse mAbs AuNPs 5 min (pre-incubation) + 15 min (LFA) 10.3 ng/mL in buffer, 9.5 ng/mL in urine, 8.0 ng/mL in serum by reader and 25 ng/mL in buffer by naked eye for B. atrox Spiked human urine, serum, & plasma 38 NPs is short for nanoparticles, AuNPs for gold nanoparticles, pAbs denote polyclonal antibodies, and mAbs denote monoclonal antibodies. a The reported detection limits are contingent on the various dilution factors used in these studies and should only be compared when considering these factors.…”

Section: Introductionmentioning

confidence: 99%

Multiplex lateral flow assay development for snake venom detection in biological matrices

Knudsen,

Belfakir,

Degnegaard

et al. 2024

Sci Rep

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Bothrops and Lachesis are two of Brazil’s medically most relevant snake genera, causing tens of thousands of bites annually. Fortunately, Brazil has good accessibility to high-quality antivenoms at the genus and inter-genus level, enabling the treatment of many of these envenomings. However, the optimal use of these treatments requires that the snake species responsible for the bite is determined. Currently, physicians use a syndromic approach to diagnose snakebite, which can be difficult for medical personnel with limited training in clinical snakebite management. In this work, we have developed a novel monoclonal antibody-based multiplex lateral flow assay for differentiating Bothrops and Lachesis venoms within 15 min. The test can be read by the naked eye or (semi)-quantitatively by a smartphone supported by a 3D-printed attachment for controlling lighting conditions. The LFA can detect Bothrops and Lachesis venoms in spiked plasma and urine matrices at concentrations spanning six orders of magnitude. The LFA has detection limits of 10–50 ng/mL in spiked plasma and urine, and 50–500 ng/mL in spiked sera, for B. atrox and L. muta venoms. This test could potentially support medical personnel in correctly diagnosing snakebite envenomings at the point-of-care in Brazil, which may help improve patient outcomes and save lives.

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“…3–4 mg of venom powder was weighed and solubilized into 200 μL 200 mM AmAc buffer (pH unaltered). As previously described, venom samples were then fractionated into 200 mM AmAc by SEC. ,, An UPLC UltiMate 3000 (Thermo Fischer Scientific, Massachusetts, United States) equipped with the Superdex Increase 200 10/300 GL column (Cytiva, Massachusetts, United States) was used in these experiments. SEC was operated at a rate of 0.5 mL/min.…”

Section: Materials and Methodsmentioning

confidence: 99%

Streamlining the Analysis of Proteins from Snake Venom

Oganesyan,

Jenkins,

Laustsen

et al. 2024

J. Proteome Res.

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Investigating snake venom is necessary for developing new treatments for envenoming and harnessing the therapeutic potential that lies within venom toxins. Despite considerable efforts in previous research, several technical challenges remain for characterizing the individual components within such complex mixtures. Here, we present native and top-down mass spectrometry (MS) workflows that enable the analysis of individual venom proteins within complex mixtures and showcase the utility of these methodologies on King cobra (Ophiophagus hannah) venom. First, we coupled ion mobility spectrometry for separation and electron capture dissociation for charge reduction to resolve highly convoluted mass spectra containing multiple proteins with masses ranging from 55 to 127 kDa. Next, we performed a top-down glycomic analysis of a 25.5 kDa toxin, showing that this protein contains a fucosylated complex glycan. Finally, temperature-controlled nanoelectrospray mass spectrometry facilitated the top-down sequence analysis of a β-cardiotoxin, which cannot be fragmented by collisional energy due to its disulfide bond pattern. The work presented here demonstrates the applicability of new and promising MS methods for snake venom analysis.

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Prototyping of a lateral flow assay based on monoclonal antibodies for detection of Bothrops venoms

Cited by 4 publications

References 31 publications

A novel lateral flow immunochromatographic assay using a recombinant VP2 antigen for total antibody detection of canine parvovirus-2

A novel lateral flow immunochromatographic assay using a recombinant VP2 antigen for total antibody detection of canine parvovirus-2

Multiplex lateral flow assay development for snake venom detection in biological matrices

Streamlining the Analysis of Proteins from Snake Venom

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