Atrazine interaction with tropical humic substances by Enzyme Linked Immunosorbent Assay

Toscano, Ilda Antonieta Salata; Gascón, Jordi; Marco, Marı́a Pilar; Rocha, Júlio César; Barceló, Damià

doi:10.1051/analusis:1998122

Cited by 16 publications

(14 citation statements)

References 15 publications

(14 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Previous work on the use of ELISA's to detect residual agrochemicals in soil samples had shown that humic substances in soil may result in an overestimation of chemical concentrations [20,27,28], and that sample dilution could be used to minimize interference [20]. As a similar type of interference might occur with biomassderived materials such as soluble lignin fragments, supernatants derived from steam pretreated corn stover (SPCS), steam pretreated poplar (SPP), steam pretreated douglas fir (SPDF), and Avicel were assessed for their possible influence on the double-antibody ELISA.…”

Section: Resultsmentioning

confidence: 99%

The Development and Use of an Elisa-Based Method to Follow the Distribution of Cellulase Monocomponents During the Hydrolysis of Pretreated Corn Stover

Pribowo¹,

Hu²,

Arantes³

et al. 2015

Fuel Production From Non-Food Biomass

View full text Add to dashboard Cite

Background: It is widely recognised that fast, effective hydrolysis of pretreated lignocellulosic substrates requires the synergistic action of multiple types of hydrolytic and some non-hydrolytic proteins. However, due to the complexity of the enzyme mixture, the enzymes interaction with and interference from the substrate and a lack of specific methods to follow the distribution of individual enzymes during hydrolysis, most of enzyme-substrate interaction studies have used purified enzymes and pure cellulose model substrates. As the enzymes present in a typical "cellulase mixture" need to work cooperatively to achieve effective hydrolysis, the action of one enzyme is likely to influence the behaviour of others. The action of the enzymes will be further influenced by the nature of the lignocellulosic substrate. Therefore, it would be beneficial if a method could be developed that allowed us to follow some of the individual enzymes present in a cellulase mixture during hydrolysis of more commercially realistic biomass substrates. Results: A high throughput immunoassay that could quantitatively and specifically follow individual cellulase enzymes during hydrolysis was developed. Using monoclonal and polyclonal antibodies (MAb and PAb, respectively), a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed to specifically quantify cellulase enzymes from Trichoderma reesei: cellobiohydrolase I (Cel7A), cellobiohydrolase II (Cel6A), and endoglucanase I (Cel7B). The interference from substrate materials present in lignocellulosic supernatants could be minimized by dilution. Conclusion: A double-antibody sandwich ELISA was able to detect and quantify individual enzymes when present in cellulase mixtures. The assay was sensitive over a range of relatively low enzyme concentration (0 -1 μg/ml), provided the enzymes were first pH adjusted and heat treated to increase their antigenicity. The immunoassay was employed to quantitatively monitor the adsorption of cellulase monocomponents, Cel7A, Cel6A, and Cel7B, that were present in both Celluclast and Accellerase 1000, during the hydrolysis of steam-pretreated corn stover (SPCS). All three enzymes exhibited different individual adsorption profiles. The specific and quantitative adsorption profiles observed with the ELISA method were in agreement with earlier work where more labour intensive enzyme assay techniques were used.

show abstract

Section: Resultsmentioning

confidence: 99%

The Development and Use of an Elisa-Based Method to Follow the Distribution of Cellulase Monocomponents During the Hydrolysis of Pretreated Corn Stover

Pribowo¹,

Hu²,

Arantes³

et al. 2015

Fuel Production From Non-Food Biomass

View full text Add to dashboard Cite

show abstract

“…To evaluate this effect, the calibration curves of BPA were prepared in 10 mM PBS containing 30 mg/L humic acid, which represented organic matters in natural waters at high level40. As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

A reusable evanescent wave immunosensor for highly sensitive detection of bisphenol A in water samples

Zhou

Liu

et al. 2014

Sci Rep

View full text Add to dashboard Cite

This paper proposed a compact and portable planar waveguide evanescent wave immunosensor (EWI) for highly sensitive detection of BPA. The incident light is coupled into the planar waveguide chip via a beveled angle through undergoing total internal reflection, where the evanescent wave field forms and excites the binding fluorophore-tagged antibodies on the chip surface. Typical calibration curves obtained for BPA has detection limits of 0.03 μg/L. Linear response for BPA ranged from 0.124 μg/L–9.60 μg/L with 50% inhibition concentration for BPA of 1.09 ± 0.25 μg/L. The regeneration of the planar optical waveguide chip allows the performance of more than 300 assay cycles within an analysis time of about 20 min for each assay cycle. By application of effective pretreatment procedure, the recoveries of BPA in real water samples gave values from 88.3% ± 8.5% to 103.7% ± 3.5%, confirming its application potential in the measurement of BPA in reality.

show abstract

“…Previous work on the use of ELISA’s to detect residual agrochemicals in soil samples had shown that humic substances in soil may result in an overestimation of chemical concentrations [20,27,28], and that sample dilution could be used to minimize interference [20]. As a similar type of interference might occur with biomass-derived materials such as soluble lignin fragments, supernatants derived from steam pretreated corn stover (SPCS), steam pretreated poplar (SPP), steam pretreated douglas fir (SPDF), and Avicel were assessed for their possible influence on the double-antibody ELISA.…”

Section: Resultsmentioning

confidence: 99%

The development and use of an ELISA-based method to follow the distribution of cellulase monocomponents during the hydrolysis of pretreated corn stover

Pribowo

Arantes

et al. 2013

Biotechnol Biofuels

View full text Add to dashboard Cite

BackgroundIt is widely recognised that fast, effective hydrolysis of pretreated lignocellulosic substrates requires the synergistic action of multiple types of hydrolytic and some non-hydrolytic proteins. However, due to the complexity of the enzyme mixture, the enzymes interaction with and interference from the substrate and a lack of specific methods to follow the distribution of individual enzymes during hydrolysis, most of enzyme-substrate interaction studies have used purified enzymes and pure cellulose model substrates. As the enzymes present in a typical “cellulase mixture” need to work cooperatively to achieve effective hydrolysis, the action of one enzyme is likely to influence the behaviour of others. The action of the enzymes will be further influenced by the nature of the lignocellulosic substrate. Therefore, it would be beneficial if a method could be developed that allowed us to follow some of the individual enzymes present in a cellulase mixture during hydrolysis of more commercially realistic biomass substrates.ResultsA high throughput immunoassay that could quantitatively and specifically follow individual cellulase enzymes during hydrolysis was developed. Using monoclonal and polyclonal antibodies (MAb and PAb, respectively), a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed to specifically quantify cellulase enzymes from Trichoderma reesei: cellobiohydrolase I (Cel7A), cellobiohydrolase II (Cel6A), and endoglucanase I (Cel7B). The interference from substrate materials present in lignocellulosic supernatants could be minimized by dilution.ConclusionA double-antibody sandwich ELISA was able to detect and quantify individual enzymes when present in cellulase mixtures. The assay was sensitive over a range of relatively low enzyme concentration (0 – 1 μg/ml), provided the enzymes were first pH adjusted and heat treated to increase their antigenicity. The immunoassay was employed to quantitatively monitor the adsorption of cellulase monocomponents, Cel7A, Cel6A, and Cel7B, that were present in both Celluclast and Accellerase 1000, during the hydrolysis of steam-pretreated corn stover (SPCS). All three enzymes exhibited different individual adsorption profiles. The specific and quantitative adsorption profiles observed with the ELISA method were in agreement with earlier work where more labour intensive enzyme assay techniques were used.

show abstract

Atrazine interaction with tropical humic substances by Enzyme Linked Immunosorbent Assay

Cited by 16 publications

References 15 publications

The Development and Use of an Elisa-Based Method to Follow the Distribution of Cellulase Monocomponents During the Hydrolysis of Pretreated Corn Stover

The Development and Use of an Elisa-Based Method to Follow the Distribution of Cellulase Monocomponents During the Hydrolysis of Pretreated Corn Stover

A reusable evanescent wave immunosensor for highly sensitive detection of bisphenol A in water samples

The development and use of an ELISA-based method to follow the distribution of cellulase monocomponents during the hydrolysis of pretreated corn stover

Contact Info

Product

Resources

About