**1. Introduction**

The global population is increasing quickly. Experts recommend that food necessities are probably going to rise significantly in the next 20 years. More than 800 million individuals,

© 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 2018 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

including 33% of the number of inhabitants in sub-Saharan Africa, are undernourished. More than 90% of them are enduring long-term malnutrition and micronutrient insufficiency. Genetic modification of crops can possibly take care of these issues. A hereditarily changed life form (GMO) is a life whose genome has been modified by methods of recombinant DNA technology. This innovation adjusts or embeds at least one quality into a life through genetic modification. GMOs hold extraordinary potential to build trim yield, enhance sustenance quality, decrease input costs and enhance creativity. To date, insect protection and herbicide resilient are the primary business attributes utilized as a part of maize, cotton and soybean [1, 2]. These qualities are giving monetary advantages to the agrochemical business, seed markets and agriculturists because of improved profitability. They additionally conceivably advantage the land because of a lessening in the utilization of chemicals or a move to the utilization of all the more naturally agreeable chemicals.

on the reaction between an antigen and a counteracting agent. Protein detection strategies for the GMO testing shift from those that are generally modest and simple to perform to more refined measures requiring costly instrumentation. Protein detection strategies can be utilized to recognize GM attributes in GM crops [6]. GMO testing has turned into a vital and essential piece of food production to ensure compliance with labeling regulations, to confirm IP frame-

Protein-Based Detection Methods for Genetically Modified Crops

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Binding assays are in widely being used in laboratories for the detection and quantification of proteins in samples. For biological samples such as urine, whole blood, plasma, serum and other biological fluids, assays are often performed in hospitals and clinical laboratories. These binding assays can likewise be performed in natural, horticultural, veterinary, mechanical athletic lawful/criminological settings and furthermore, snappy discovery of irresistible sicknesses in serological testing of people and creatures. The principles involved in such assays are well known by those skilled in the art. Many such devices have been described and are available commercially. Immunological binding assay is the sandwich assay. However, in clinical laboratories, the use of solid phase chromatographic binding assay devices has become commonplace for their relative ease of use, economy, and reproducibility. Typically, these chromatographic assay devices are comprised of a porous chromatographic medium which acts as the matrix for the binding assay. The sample is added directly or indirectly to one end of the medium and is chromatographically transported to a detection reagent with which it reacts to form a labeled product, which is then transported to a test zone containing an immobilized capture reagent such as a capture antibody, in which the presence, absence or

This depends on immunochromatography and sidelong stream measures. It identifies with immunomeasure dipsticks, and especially to those test gadgets used to lead immunological and serological restricting tests. This new strip test is low in cost, quick, monetary, convenient and less laborious. It can be utilized to detect qualitatively or semiquantitatively the presence of protein of interest samples. The development in techniques for utilizing such dipsticks for

GMOs hold the awesome potential to increase crop yield, enhance nutrient quality, lessen input costs and enhance creativity. To date, insect and herbicide resistance are the principal business attributes utilized in maize, cotton and soybean. The worldwide area of GM crops expanded 47-fold, from 1.7 million hectares in 1996 to 81 million hectares in 2004, with an expanding extent developed by developing nations [2, 3]. GMO testing has turned into an indispensable and essential piece of food production to ensure compliance with labeling regulations. Protein-based methods, for example, ELISA and strip tests are viable for natural items yet rely upon the accessibility of business units and are not appropriate for prepared items because of protein degradation [7]. Parallel stream systems are subjective or semiquantitative [4]. Immunoassays have the ability to be broadly executed on a large scale for the recognition of novel proteins in crude food items. Immunoassay advances are perfect for the subjective and quantitative discovery of many sorts of proteins and pathogens in complex systems [5, 8]. Effective testing of GM products must be accomplished with the improvement

works and secure customers by approving "non-GMO" item publicizing claims [7].

quantity of an analyte can be determined.

the detection of GMOs is increasing.

The development of GM crops is progressing, more qualities are rising and a bigger number of sections of land are being planted with GM crops. The arrival of GM harvests and items in the business sectors worldwide has expanded the administrative need to screen and check the nearness and the measure of GM crops in yields. The worldwide region of GM crops expanded from 1.7 million hectares in 1996 to 81 million hectares in 2004, with an expanding extent developed by creating nations. More than 8 million ranchers are profiting from this innovation [3]. Around 90% of the farmers are small agriculturists from developing nations, who increased their earnings from biotech crops significantly.

The administrative need to screen and confirm the nearness and the measure of GM crops has expanded with the development of the GM crops [4]. Effective monitoring of GM crops must be accomplished with the improvement of proper techniques.

GM crops can be distinguished by identifying the transformed hereditary material at the DNA level, the subsequent protein or phenotype. A few expository techniques, for example, strategies in view of the polymerase chain reaction (PCR) for identifying the incorporated DNA, immunological measures for detecting the subsequent protein or utilizing bioassays to recognize the resultant phenotype have been reported. Western blotting, enzyme-linked immunosorbent assay (ELISA) and parallel stream sticks are common protein-based test techniques [5]. A few other diagnostic advances that can give answers for current specialized issues in the GM test examinations are rising. These techniques incorporate mass spectrometry, chromatography, close infrared spectroscopy, miniaturized scale manufactured gadgets and, specifically, DNA chip innovation (microarrays) and mostly immunoassays. Different immunoassays are being used to determine the genetically modified proteins.

The test on a specimen is, for the most part, a screening test that may distinguish a scope of GMOs. This can be trailed by a particular test to recognize the type of GMO in the sample and additionally intended to measure the quantity of a particular GMO. The lion's share of protein recognition strategies depends on immunoassays for discovery and evaluation of new (outside) proteins presented through the genetic modification of plants. Immunoassay depends on the reaction between an antigen and a counteracting agent. Protein detection strategies for the GMO testing shift from those that are generally modest and simple to perform to more refined measures requiring costly instrumentation. Protein detection strategies can be utilized to recognize GM attributes in GM crops [6]. GMO testing has turned into a vital and essential piece of food production to ensure compliance with labeling regulations, to confirm IP frameworks and secure customers by approving "non-GMO" item publicizing claims [7].

including 33% of the number of inhabitants in sub-Saharan Africa, are undernourished. More than 90% of them are enduring long-term malnutrition and micronutrient insufficiency. Genetic modification of crops can possibly take care of these issues. A hereditarily changed life form (GMO) is a life whose genome has been modified by methods of recombinant DNA technology. This innovation adjusts or embeds at least one quality into a life through genetic modification. GMOs hold extraordinary potential to build trim yield, enhance sustenance quality, decrease input costs and enhance creativity. To date, insect protection and herbicide resilient are the primary business attributes utilized as a part of maize, cotton and soybean [1, 2]. These qualities are giving monetary advantages to the agrochemical business, seed markets and agriculturists because of improved profitability. They additionally conceivably advantage the land because of a lessening in the utilization of chemicals or a move to the utilization of all the more naturally agreeable

The development of GM crops is progressing, more qualities are rising and a bigger number of sections of land are being planted with GM crops. The arrival of GM harvests and items in the business sectors worldwide has expanded the administrative need to screen and check the nearness and the measure of GM crops in yields. The worldwide region of GM crops expanded from 1.7 million hectares in 1996 to 81 million hectares in 2004, with an expanding extent developed by creating nations. More than 8 million ranchers are profiting from this innovation [3]. Around 90% of the farmers are small agriculturists from developing nations,

The administrative need to screen and confirm the nearness and the measure of GM crops has expanded with the development of the GM crops [4]. Effective monitoring of GM crops must

GM crops can be distinguished by identifying the transformed hereditary material at the DNA level, the subsequent protein or phenotype. A few expository techniques, for example, strategies in view of the polymerase chain reaction (PCR) for identifying the incorporated DNA, immunological measures for detecting the subsequent protein or utilizing bioassays to recognize the resultant phenotype have been reported. Western blotting, enzyme-linked immunosorbent assay (ELISA) and parallel stream sticks are common protein-based test techniques [5]. A few other diagnostic advances that can give answers for current specialized issues in the GM test examinations are rising. These techniques incorporate mass spectrometry, chromatography, close infrared spectroscopy, miniaturized scale manufactured gadgets and, specifically, DNA chip innovation (microarrays) and mostly immunoassays. Different immunoassays are being used to determine the geneti-

The test on a specimen is, for the most part, a screening test that may distinguish a scope of GMOs. This can be trailed by a particular test to recognize the type of GMO in the sample and additionally intended to measure the quantity of a particular GMO. The lion's share of protein recognition strategies depends on immunoassays for discovery and evaluation of new (outside) proteins presented through the genetic modification of plants. Immunoassay depends

who increased their earnings from biotech crops significantly.

be accomplished with the improvement of proper techniques.

chemicals.

48 Protein-Protein Interaction Assays

cally modified proteins.

Binding assays are in widely being used in laboratories for the detection and quantification of proteins in samples. For biological samples such as urine, whole blood, plasma, serum and other biological fluids, assays are often performed in hospitals and clinical laboratories. These binding assays can likewise be performed in natural, horticultural, veterinary, mechanical athletic lawful/criminological settings and furthermore, snappy discovery of irresistible sicknesses in serological testing of people and creatures. The principles involved in such assays are well known by those skilled in the art. Many such devices have been described and are available commercially. Immunological binding assay is the sandwich assay. However, in clinical laboratories, the use of solid phase chromatographic binding assay devices has become commonplace for their relative ease of use, economy, and reproducibility. Typically, these chromatographic assay devices are comprised of a porous chromatographic medium which acts as the matrix for the binding assay. The sample is added directly or indirectly to one end of the medium and is chromatographically transported to a detection reagent with which it reacts to form a labeled product, which is then transported to a test zone containing an immobilized capture reagent such as a capture antibody, in which the presence, absence or quantity of an analyte can be determined.

This depends on immunochromatography and sidelong stream measures. It identifies with immunomeasure dipsticks, and especially to those test gadgets used to lead immunological and serological restricting tests. This new strip test is low in cost, quick, monetary, convenient and less laborious. It can be utilized to detect qualitatively or semiquantitatively the presence of protein of interest samples. The development in techniques for utilizing such dipsticks for the detection of GMOs is increasing.

GMOs hold the awesome potential to increase crop yield, enhance nutrient quality, lessen input costs and enhance creativity. To date, insect and herbicide resistance are the principal business attributes utilized in maize, cotton and soybean. The worldwide area of GM crops expanded 47-fold, from 1.7 million hectares in 1996 to 81 million hectares in 2004, with an expanding extent developed by developing nations [2, 3]. GMO testing has turned into an indispensable and essential piece of food production to ensure compliance with labeling regulations. Protein-based methods, for example, ELISA and strip tests are viable for natural items yet rely upon the accessibility of business units and are not appropriate for prepared items because of protein degradation [7]. Parallel stream systems are subjective or semiquantitative [4]. Immunoassays have the ability to be broadly executed on a large scale for the recognition of novel proteins in crude food items. Immunoassay advances are perfect for the subjective and quantitative discovery of many sorts of proteins and pathogens in complex systems [5, 8]. Effective testing of GM products must be accomplished with the improvement of proper strategies for detection. These strategies are for the most part in view of the study of the novel proteins or DNA.

**3. Classification of immunoassays**

**1.** Competitive homogenous immunoassays **2.** Competitive heterogeneous immunoassays

**3.** One-site noncompetitive immunoassays **4.** Two-site noncompetitive immunoassays

unbound analyte in the sample.

sample.

**3.1. Competitive, homogeneous immunoassays**

**3.2. Competitive heterogeneous immunoassay**

remaining labeled and bound analyte is measured.

**3.3. One-site noncompetitive immunoassays**

**3.4. Two-site noncompetitive immunoassays**

is measured. The details of the labels are given as follows:

In competitive homogenous immunoassay, there is a competition between labeled and bound analyte (bound to the antibody) with unlabeled and unbound analyte in the sample. As a competition, the unlabeled and unbound analyte displace the labeled and bound analyte and get them attached in place, while the detached labeled analyte then give fluorescence, and this fluorescence is measured, which is proportional to the amount of unlabeled and initial

Protein-Based Detection Methods for Genetically Modified Crops

http://dx.doi.org/10.5772/intechopen.75520

51

In heterogeneous assay, there is a competition between bound and labeled analyte (bound to the antibody) with unbound and unlabeled analyte, the difference from competitive homogenous assay is that the labeled unbound/displaced analyte is separated by washing, and the

In this immunoassay, the amount of unknown analyte in the sample is measured by adding the labeled antibodies. The labeled antibodies get attached with the analyte in the sample, and the extra labeled unbound antibodies are washed away, so, only labeled and bound antibodies are present in the sample, the intensity of fluorescence of these antibodies is measured, which is proportional to the 3.4. amount of unknown analyte in the

In this immunoassay, there is an antibody present on a site, and the analyte in a sample is added to the antibody get attached, and then second antibody is added which is attached with the label. If the specific analyte is not present in the solution, the second antibody will not attach. Then, the fluorescence of the labeled antibody is measured, which is directly proportional to the amount of analyte in the sample. It is very important to consider that there are washing steps after every reaction, so extra materials are always washed away. The other thing very important is that what type of labels are attached and how the fluorescence/signal

For approval of a scientific strategy, the testing objective must be characterized and execution qualities must be illustrated. Execution qualities incorporate exactness, extraction proficiency, accuracy, reproducibility, affectability, specificity and strength. The utilization of approved strategies is essential to guarantee acknowledgment of results delivered by diagnostic research facilities [9]. The greater part of protein detection techniques depends on immunoassays. Protein detection techniques can possibly recognize the nearness of a particular GM quality and to give the total measurement of the level of transgene expression. Protein identification strategies are exceedingly reasonable for checking particular GM attributes amid treatment of crude items, gave the protein is communicated in the piece of the plant being tried.

Here are the details of immunoassays being used for the detection of genetically modified proteins.
