Polyclonal-Antibody-Based ELISA To Detect Milk Alkaline Phosphatase

Vega‐Warner, A. V.; Gandhi, Hasand; Smith, Denise M.; Ustunol, Z.

doi:10.1021/jf990412v

Cited by 14 publications

(14 citation statements)

References 12 publications

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“…A large number of reports regarding the development of enzyme immunoassays using peroxidase as reporter enzyme to detect toxins, pathogens, and other analyses are available. For example, in detection of Goynyautoxins, Chlamydia, Fusarium toxin, Dengue virus proteins, Hepatitis-E virus peroxidases are used [194][195][196][197][198][199]. HRP-based immunoassays for the detection of undeclared milk proteins (eg.…”

Section: Analytical Applicationsmentioning

confidence: 99%

A Comprehensive Review on Function and Application of Plant Peroxidases

Pandey¹,

Awasthi²,

Singh³

et al. 2017

Biochem Anal Biochem

276

213

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Peroxidases, one of the key antioxidant enzymes, are widely distributed in nature and catalyze oxidation of various electron donor substrates concomitant with the decomposition of H 2 O 2. The non-animal plant peroxidases (class III peroxidase) are involved in various essential physiological processes of plant growth and development throughout their life cycle. In view of the capability of peroxidases to catalyze the redox reaction for a wide range of substrates, they are considered as one of the important enzyme from the point of view of their various medicinal, biochemical, immunological, biotechnological and industrial applications. They have been successfully used for biopulping and biobleaching in the paper and textile industries. Peroxidases have also been used in organic synthesis, bioremediation, as well as various analytical applications in diagnostic kits, ELISA. Peroxidase based biosensors find application in analytical systems for determination of hydrogen peroxide, glucose, alcohols, glutamate, and choline etc. Thus, in view of array of physiological functions as well as industrial applications, the peroxidases have conquered a dominant position in research groups and become one of the most extensively studied enzymes. In this direction, the present review embodies the classification, mechanism of action, major physiological functions as well as industrial applications of plant peroxidases.

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Section: Analytical Applicationsmentioning

confidence: 99%

A Comprehensive Review on Function and Application of Plant Peroxidases

Pandey¹,

Awasthi²,

Singh³

et al. 2017

Biochem Anal Biochem

276

213

View full text Add to dashboard Cite

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“…Detecting LF at low concentrations is relevant in the context of bioterrorism since Bacillus anthracis is classified as a biowarfare agent 8-12 and since sensitive methods for detection of LF are important for early determination of infection by B. anthracis 13. Although colorimetric,14-17 chromatographic,18-20 electrochemical,21-23 and ELISA-based 24, 25 methods have been developed to quantify the activity of clinically relevant enzymes, new diagnostic platforms are still needed to improve the robustness, sensitivity, selectivity, portability, speed, and cost-effectiveness of bioanalytical assays.…”

mentioning

confidence: 99%

A Semi-Synthetic Ion Channel Platform for Detection of Phosphatase and Protease Activity

et al. 2009

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Sensitive methods to probe the activity of enzymes are important for clinical assays and for elucidating the role of these proteins in complex biochemical networks. This paper describes a semisynthetic ion channel platform for detecting the activity of two different classes of enzymes with high sensitivity. In the first case, this method uses single ion channel conductance measurements to follow the enzyme-catalyzed hydrolysis of a phosphate group attached to the C-terminus of gramicidin A (gA, an ion channel-forming peptide) in the presence of alkaline phosphatase (AP). Enzymatic hydrolysis of this phosphate group removes negative charges from the entrance of the gA pore, resulting in a product with measurably reduced single ion channel conductance compared to the original gA-phosphate substrate. This technique employs a standard, commercial bilayer setup and takes advantage of the catalytic turnover of enzymes and the amplification characteristics of ion flux through individual gA pores to detect picomolar concentrations of active AP in solution. Furthermore, this technique makes it possible to study the kinetics of an enzyme and provides an estimate for the observed rate constant (k cat ) and the Michaelis constant (K M ) by following the conversion of the gA-phosphate substrate to product over time in the presence of different concentrations of AP. In the second case, modification of gA with a substrate for proteolytic cleavage by anthrax lethal factor (LF) afforded a sensitive method for detection of LF activity, illustrating the utility of ion channel-based sensing for detection of a potential biowarfare agent. This ion channelbased platform represents a powerful, novel approach to monitor the activity of femtomoles to picomoles of two different classes of enzymes in solution. Furthermore, this platform has the potential for realizing miniaturized, cost-effective bioanalytical assays that complement currently established assays. Keywords

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“…However, the presence of ALP activity in properly pasteurized milk as a consequence of contamination with bacterial ALPs has been reported (Hammer & Olson, 1941). To quantify milk ALP without interference by non-bovine milk ALPs, ELISA tests based on polyclonal antibodies against purified bovine milk ALP have been proposed (Vega-Warner et al 2000; Chen et al 2006). However, none of them has been proven to quantify dilutions of raw milk in pasteurized milk.…”

mentioning

confidence: 99%

Inactivation-denaturation kinetics of bovine milk alkaline phosphatase during mild heating as determined by using a monoclonal antibody-based immunoassay

Levieux¹,

Geneix²,

Levieux³

2007

Journal of Dairy Research

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A monoclonal antibody based capture immunoassay has been recently developed for the specific quantitation of bovine milk alkaline phosphatase (ALP) without interference by contaminating microbial or fungal ALPs (Geneix et al. 2007). This immunoassay was used to study the kinetics of ALP heat denaturation in bovine milk over a range 50-60 degrees C for 5 to 60 min using a colorimetric quantification of the enzyme activity as a reference test. A denaturation midpoint was obtained at 56 degrees C for a 30 min heating. Thermal inactivation was found to follow first order kinetics and is characterized by z value of 6.7 deg C (D60 degrees C=24.6 min) and 6.8 (D60 degrees C=23.0 min) for respectively immunoassay and colorimetric assay. The high values of enthalpy of activation and the positive values of the entropy of activation and free energy of activation indicate that during denaturation ALP underwent a large change in conformation. The results of the immunoassay were highly correlated (r=0.994) with those obtained by the colorimetric assay. A similar high correlation (r=0.998) was obtained when industrially thermized milks (62-67 degrees C for 20-90 s) were analysed by both techniques. These results indicated that 1) thermally induced epitopic structural changes recognized by the capture monoclonal antibody are concomitant with or occur after the loss of enzymatic activity and 2) quantification of ALP by the specific immunoassay is appropriate for determining mild time/temperature treatment of milk and for the control of milk pasteurization.

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Polyclonal-Antibody-Based ELISA To Detect Milk Alkaline Phosphatase

Cited by 14 publications

References 12 publications

A Comprehensive Review on Function and Application of Plant Peroxidases

A Comprehensive Review on Function and Application of Plant Peroxidases

A Semi-Synthetic Ion Channel Platform for Detection of Phosphatase and Protease Activity

Inactivation-denaturation kinetics of bovine milk alkaline phosphatase during mild heating as determined by using a monoclonal antibody-based immunoassay

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