الفهرس 
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Abstract
A new food crop or food ingredient derived from a genetically engineered (GE) organism must undergo safety assessment including allergenicity assessment prior to its release into the market. This thesis included three studies using literature review of the history of safe use of transgenic proteins, the gene donor, and the gene recipient. The first study included comparison of IgE binding from individual soybean allergic subjects to proteins in a new GE soybean line and other commercial line. To maintain the aim of this study the following was carried out: Protein extraction from a genetically modified soybean (roundup soybean) and from 8 non-genetically modified soybean samples from diverse sources representing different environments, also from corn and peanut, Electrophoresis of protein extracts using SDS PAGE, Use of immunoblotting technique to transfer proteins to a membrane (Polyvinylidene difluoride) and sensitization using sera from soybean allergic sera, ELISA (enzyme-linked immunosorbent assay.
The results indicated that the GE soybean line did not present an increased risk for soybean allergic subjects especially because those with soybean allergy should avoid all soybeans.
The second study evaluated the cross reactivity between soybean and peanut major allergens. To achieve the aim of this study the following was carried out: Preparation of glycinin and β-conglycinin from extracts of soybean, Detecting glycoproteins, HiTrap™ Con A 4B Purification, Batch Anion exchange purification, Quantification using liquid chromatography-tandem mass spectrometry (LC-MS/MS), SDS-PAGE and immunoblotting, Inhibition immunoblotting, Mediator release assay.
The results showed that cross-reactive IgE binding between soybean and peanut was common but not constant among the study population. The subjects used in this study showed specific binding to peanut and soybean proteins. The third study evaluated the risk of a recombinant purified protein of Acetohydroxy Acid Synthase purified from camelina line 14CS851-01-14. Assay parameters used in this study included verification of pepsin activity, established limit of detection of the protein in the stained gel (at 10% total stainable protein) and use of an objective measurement of the time of digestion required to reach 90% digestion. The activity of the pepsin in SGF was tested on each day of assay based on digestion of bovine hemoglobin to ensure that it is within a tolerance interval reported by Worthington for that lot of enzyme. This protein underwent in vitro pepsin digestion to evaluate its stability. The digestion was performed at 37°C and samples are removed at specific times and the activity of pepsin is quenched by neutralization with carbonate buffer and Laemmli loading buffer, then heating to more than 85°C for 10 minutes. The timed digestion samples are separated by SDS-PAGE and stained with Coomassie blue to evaluate the extent of digestion. The results of this study demonstrated that the intact AHAS protein supplied by Linnaeus Plant Sciences, Inc. is completely digested in pepsin at pH 2.0 at 10 units (as per standard protocol) and 1 µg AHAS after 60 minutes. No residual undigested bands were visible after 0.5 minutes.