Development and Validation of an Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Lysozyme in Cheese

Schneider, Nadine; Weigel, Ingrid; Werkmeister, K.; Pischetsrieder, Monika

doi:10.1021/jf9025019

Cited by 48 publications

(14 citation statements)

References 28 publications

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“…Peng et al reported a faradic impedance spectroscopy aptasensor using aptamer as recognition element for lysozyme analysis ). Schneider et al developed an enzyme-linked immunosorbent assay using antibody as recognition element (Schneider et al, 2010). However, the linear ranges of these methods were narrower than that of this article and recognition elements of these methods were low stability and high cost.…”

Section: Samplesmentioning

confidence: 85%

Magnetic molecularly imprinted nanoparticles for recognition of lysozyme

Dai

et al. 2010

Biosensors and Bioelectronics

168

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Section: Samplesmentioning

confidence: 85%

Magnetic molecularly imprinted nanoparticles for recognition of lysozyme

Dai

et al. 2010

Biosensors and Bioelectronics

168

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“…However, these methods require pre-treatment of the sample, expensive tools, qualified personnel and long-time analysis. The enzyme-linked immunosorbent assay (ELISA) is also used to this aim [32,35], but this approach has several disadvantages such as the nonspecific adsorption of substances generating false positives/negatives [36], the possible matrix effect on enzyme activity [37] and kit high cost. To overcome these problems, a multitude of sensors for lysozyme have been developed in recent years.…”

Section: Introductionmentioning

confidence: 99%

Molecularly Imprinted Polyscopoletin for the Electrochemical Detection of the Chronic Disease Marker Lysozyme

2020

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Herein we report the electropolymerization of a scopoletin based molecularly imprinted polymer (MIP) for the detection of lysozyme (Lyz), an enzymatic marker of several diseases in mammalian species. Two different approaches have been used for the imprinting of lysozyme based, respectively, on the use of a monomer-template mixture and on the covalent immobilization of the enzyme prior to polymer synthesis. In the latter case, a multi-step protocol has been exploited with preliminary functionalization of gold electrode with amino groups, via 4-aminothiophenol, followed by reaction with glutaraldehyde, to provide a suitable linker for lysozyme. Each step of surface electrode modification has been followed by cyclic voltammetry and electrochemical impedance spectroscopy, which has been also employed to test the electrochemical responses of the developed MIP. The sensors show good selectivity to Lyz and detect the enzyme at concentrations up to 292 mg/L (20 μM), but with different performances, depending on the used imprinting approach. An imprinting factor equal to 7.1 and 2.5 and a limit of detection of 0.9 mg/L (62 nM) and 2.1 mg/L (141 nM) have been estimated for MIPs prepared with and without enzyme immobilization, respectively. Competitive rebinding experiment results show that this sensing material is selective for Lyz determination. Tests were performed using synthetic saliva to evaluate the potential application of the sensors in real matrices for clinical purposes.

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“…In Italy, the use of lysozyme is widespread (Panari and Filippi, 2009); lysozyme has been detected in Grana Padano, in grated hard cheese mixtures (Cocolin et al, 2004;Iaconelli et al, 2008), and in semihard goat and ewe cheeses (Dragoni et al, 2011;Schneider et al, 2010bSchneider et al, , 2011. Even though the current commercial source for lysozyme is from HEW, it is present also in milk, blood serum, tears, and saliva (Callewaert and Michiels, 2010).…”

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confidence: 99%

Short communication: Jenny milk as an inhibitor of late blowing in cheese: A preliminary report

Cosentino

Paolino

Freschi

et al. 2013

Journal of Dairy Science

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Late blowing on semihard and hard cheese may have an important economic effect on dairy production. Many studies have attempted to prevent this defect by physical treatment, the use of additives, and the use of bacteriocins. In this paper, we look at the effect of jenny milk as an inhibitor of blowing caused by clostridia and coliforms in ewe cheese making. Bulk ewe and jenny milk samples were collected in the morning by mechanical milking and were refrigerated at 4°C. On the collected samples, the count of somatic cells, coliforms, Clostridium butyricum, and Escherichia coli were determined. The bulk raw milk was divided in two 45-L vats: vat 1 was used as a control, whereas 0.5L of jenny milk was added to vat 2. Four semihard cheeses, weighing about 2 kg each, were made from each vat. Cheese making was replicated twice. After a ripening period of 60 d, the count of coliforms and of C. butyricum was determined. In the treated group, a significant inhibition of coliform bacteria was observed. The addition of jenny milk in cheese making may prove to be a useful and innovative approach for the inhibition of spore-forming clostridia strains.

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Development and Validation of an Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Lysozyme in Cheese

Cited by 48 publications

References 28 publications

Magnetic molecularly imprinted nanoparticles for recognition of lysozyme

Magnetic molecularly imprinted nanoparticles for recognition of lysozyme

Molecularly Imprinted Polyscopoletin for the Electrochemical Detection of the Chronic Disease Marker Lysozyme

Short communication: Jenny milk as an inhibitor of late blowing in cheese: A preliminary report

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