Validation of an extraction method for Cry1Ab protein from soil

Mueting, Sara A.; Strain, Katherine E.; Lydy, Michael J.

doi:10.1002/etc.2383

Cited by 10 publications

(6 citation statements)

References 34 publications

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“…The one difference that might explain the decline of Bt proteins from the water in their experiment is the presence of sediment in their microcosm, whereas sediment was absent from our experimental set-up. It is known that Cry proteins strongly adsorb to surface active particles of clay and organic matter in soils (Stotzky, 2005;Mueting et al, 2014), but in the absence of sediment, the Cry protein concentration increased up until the termination of our experiment at the end of Day 16.…”

Section: Resultsmentioning

confidence: 69%

Short communication: The rate of release of Cry1Ab protein from Bt maize leaves into water

Pisanie

Preez

Berg

et al. 2019

WSA

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Transgenic Bt maize plants are genetically modified to contain genes of Bacillus thuringiensis that encode for δ-endotoxins (Cry1Ab protein) that have insecticidal properties. These endotoxins target certain lepidopteran pests of maize. There are several entry routes by which Cry proteins enter the aquatic ecosystem in which aquatic organisms are exposed to these proteins. The main route is through plant debris such as leaves, stalks and postharvest detritus that are transported by means of run-off, rain and wind. While several studies have been conducted on the fate of Cry1Ab protein in terrestrial ecosystems, little is known of the release rates of Cry1Ab proteins from maize plant tissues that end up in aquatic ecosystems. In this study, leaves of Bt-maize and its isoline were submerged in containers filled with deionised or borehole water for a period of 16 days, and kept at 3 different temperatures (10±1, 21±1 and 30±1°C). Samples were collected at 1, 2, 4, 8, 16, 24, 48, 96, 192 and 384 h post submersion and analysed for Cry protein content using enzyme-linked immunosorbent assay (ELISA). The release of Cry protein from submerged maize leaves was influenced by temperature, and duration of immersion. An increase in Cry protein levels in the water was observed from the first hour onwards in both water types until the end of the experiment. The highest concentration of Cry protein was found at 30°C. This study showed that temperature and time period influence the release rate of Cry proteins from dried leaf matter into the aquatic environment.

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Section: Resultsmentioning

confidence: 69%

Short communication: The rate of release of Cry1Ab protein from Bt maize leaves into water

Pisanie

Preez

Berg

et al. 2019

WSA

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“…() noted that adsorption of Cry1Ab decreased with increasing organic carbon content. Both increases and decreases in adsorption of Cry proteins, however, have been observed after chemical removal of soil organic matter (Crecchio & Stotzky, ; Muchaonyerwa et al ., , ; Mueting et al ., ). When the ratio of organic matter to clay is small, most organic matter will be in the form of clay–organic matter complexes (Dexter et al ., ), and organic coatings might compete with protein for adsorption.…”

Section: Resultsmentioning

confidence: 99%

“…Various extraction cocktails have been proposed and tested. The greatest extraction efficiency is found, as for other soil proteins including enzymes, when extraction solutions have an alkaline pH and contain surfactants, a large salt concentration and a competitive protein (Palm et al, 1994;Fornasier & Margon, 2007;Helassa et al, 2011;Mueting et al, 2014). The role of the competitive protein, in this case bovine serum albumin (BSA), is to replace and prevent readsorption of the Cry proteins on soil surfaces by a mechanism of heteromolecular exchange.…”

Section: Introductionmentioning

confidence: 99%

Comparison of the affinity and extraction yield of trace amounts of three Cry proteins from Bacillus thuringiensis in contrasting types of soil

Hung

Truong

Binh

et al. 2015

European J Soil Science

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Summary The use of insecticidal proteins known as Cry or Bt, either as biopesticides used in agriculture or as vector control or originating from commercial genetically modified crops (GM), is increasing rapidly. The fate of these proteins in the environment depends strongly on their adsorption on the organo–mineral complexes of soil. Environmental monitoring requires the quantification of the proteins and this entails their chemical extraction from soil. Three Cry proteins, Cry1Ac, Cry1C and Cry2A, present in commercial biopesticide formulations or synthesized by GM plants or both were studied. The adsorption of trace amounts of Cry proteins on over 40 types of soil with contrasting properties was measured in dilute suspension. After a short incubation the extraction yield was measured with a previously tested alkaline solution that contained surfactant and another protein. Each of the proteins had a strong affinity for soil. No soil property was observed to determine either the affinity for soil or the extraction yield. There was no simple relation between the affinity (assessed from the distribution coefficient, Kd) and the extraction yield, although there was a significant inverse relation (P < 0.05) for two of the proteins, Cry1Ac and Cry2A. The proteins differ in both their affinity for soil and their extraction yields. We conclude that these insecticidal proteins will be largely immobile in soil, but that routine environmental monitoring can give only semi‐quantitative values for protein in soil.

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“…These criteria, as well as the scope of the validation procedure, may vary according to the intended use of the assay; however, at a minimum, validation procedures typically address the sensitivity, specificity, accuracy, and precision of ELISAs to ensure that the target analyte can be detected and/or quantified in a reproducible manner. Several excellent articles and book chapters have been published on the topic of ELISA validation and cover the topic in greater detail than in this review. ,, Additionally, several articles have been published describing validated methods for detection of Cry proteins in various matrices. ,− The remainder of this section will briefly cover the key areas of validation: sensitivity, specificity, accuracy, and precision.…”

Section: Validation Of Elisasmentioning

confidence: 99%

A Review of Cry Protein Detection with Enzyme-Linked Immunosorbent Assays

Albright

Hellmich

Coats

2016

J. Agric. Food Chem.

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The widespread use of Cry proteins in insecticide formulations and transgenic crops for insect control has led to an increased interest in the environmental fate of these proteins. Although several detection methods are available to monitor the fate of Cry proteins in the environment, enzyme-linked immunosorbent assays (ELISAs) have emerged as the preferred detection method, due to their cost-effectiveness, ease of use, and rapid results. Validation of ELISAs is necessary to ensure accurate measurements of Cry protein concentrations in the environment. Validation methodology has been extensively researched and published for the areas of sensitivity, specificity, accuracy, and precision; however, cross validation of ELISA results has been studied to a lesser extent. This review discusses the use of ELISAs for detection of Cry proteins in environmental samples and validation of ELISAs and introduces cross validation. The state of Cry protein environmental fate research is considered through a critical review of published literature to identify areas where the use of validation protocols can be improved.

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Validation of an extraction method for Cry1Ab protein from soil

Cited by 10 publications

References 34 publications

Short communication: The rate of release of Cry1Ab protein from Bt maize leaves into water

Short communication: The rate of release of Cry1Ab protein from Bt maize leaves into water

Comparison of the affinity and extraction yield of trace amounts of three Cry proteins from Bacillus thuringiensis in contrasting types of soil

A Review of Cry Protein Detection with Enzyme-Linked Immunosorbent Assays

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